Influential factors for optimizing and strengthening mesenchymal stem cells and hematopoietic stem cells co-culture

doi:10.21203/rs.3.rs-2191628/v1

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Influential factors for optimizing and strengthening mesenchymal stem cells and hematopoietic stem cells co-culture

https://doi.org/10.21203/rs.3.rs-2191628/v1

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published 25 Jan, 2024

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Mesenchymal stem cells [MSCs] and Hematopoietic stem cells [HSCs] are two types of bone marrow stem cells that could proliferate and differentiate into different cell lineages. HSCs interact with MSCs under the protective conditions, called niche. Numerous studies have indicated supportive effects of MSCs on HSCs proliferation and differentiation. Furthermore, HSCs have many clinical applications and could treat different hematologic and non-hematologic diseases. For this purpose, there is a need to perform in vitro studies to optimize their expansion. Therefore, various methods including co-culture with MSCs are used to address the limitations of HSCs culture. Some parameters that might be effective for improving the co-culture system, such as MSC paracrine profile, scaffolds, hypoxia, culture medium additives, and the use of various MSC sources, have been examined in different studies. In this article, we investigated the potential factors for optimizing HSCs/ MSCs co-culture. It might be helpful to apply a suitable approach for providing high quality HSCs and improving their therapeutic applications in the required fields.

Mesenchymal stem cell

Hematopoietic stem cells

Co-culture

Hypoxia.

Stem cells are primitive cells with at least two characteristics, including self-renewal and differentiation into various lineages [1]. There are two main populations of stem cells in the bone marrow, the hematopoietic stem cells (HSCs) and the mesenchymal stromal cells (MSCs) [2]. MSCs are valuable cells that could support other bone marrow cells by their close relationship with them [1]. These cells also play an essential role in the proliferation, survival, and differentiation of HSCs through the secretion of growth factors and cell-cell contact. Moreover, MSCs can be used as a feeder layer in HSCs culture [3].

MSCs are plastic-adhesive spindle-shaped cells that expanded in specific culture media in vitro and are one of the most critical supporting cells for the hematopoiesis system in vivo [4, 1]. These cells express CD73, CD90, CD105, CD16, and HLA-ABC and do not express CD31, CD34, CD45, CD80, and HLA-DR [1, 5, 6]. MSCs are progenitor cells that can differentiate into different cells under specific conditions [6, 7].

MSCs can be isolated from different tissues, including adipose tissue, amniotic fluid [6], bone marrow, liver, lung, spleen [8], placenta, umbilical cord blood [9], Wharton jelly, and other sources. MSC secretes cytokines, chemokines, and growth factors [10]. These properties can be utilized in the disease treatment and regenerative medicine [6, 10]. The MSC-secreted cytokines, including Interleukin 6 (IL-6), FMS-like tyrosine kinase 3 ligand (FLT3L), stem cell factor (SCF), granulocyte colony-stimulating factor (G-CSF), and granulocyte monocyte colony-stimulating factor (GM-CSF), involve in HSCs differentiation and proliferation [11].

MSCs are intrinsically capable of secreting other cytokines such as Thrombopoietin (TPO), Angiopoietins (Angs), and wingless-related integration (Wnt), which involve in the survival, quiescence, and self-renewal of HSCs [1]. MSCs produce growth factors that influence the maintenance of HSCs´ critical features in the culture media. These cells additionally provide proper cell-cell contact and extracellular binding to support HSCs expansion [12].

Different blood cells differentiate from HSCs [13]. These mentioned cells produce common myeloid and common lymphoid progenitor cells [14]. HSCs could be divided into two categories based on their self-renewal ability: long-term hematopoietic stem cells (LT-HSCs) and short-term hematopoietic stem cells (ST-HSCs) [13].

LT-HSCs have high self-renewal capacity and are less affected by cytokines. ST-HSCs have low self-renewal capacity, are more active, and induce hemopoiesis for less time in vitro [13].

HSCs have markers, including CD34, expressed on all HSCs, and CD90; expressed on the primitive HSCs. On the other hand, the lack of CD38 marker on these cells indicates the early stages of hematopoiesis. Differentiation of HSCs into lymphoid and myeloid cells lead to expression of specific markers of each lineage [15].

HSCs status is affected by a specific microenvironment, called a niche, in the bone marrow. HSCs niche affects their fate by regulating their self-renewal, proliferation, and differentiation [15–17]. The HSCs niche is composed of various cells such as MSCs, osteoblasts, adipocytes, and fibroblasts [16].

HSCs are expanded in the laboratory using various methods for application in the therapeutic or research fields. Simulation of HSCs niche and utilization of MSC in HSC ex vivo co-culture, promotes HSC culturing condition and provides an adequate number of HSCs [18]

MSCs enhance the survival and self-renewal of HSCs by either secreting cytokines and growth factors or mediating direct cell-cell contact via adhesion molecules [2]. In other words, co-culturing of HSCs with MSCs mimics the natural hematopoietic microenvironment for HSC proliferation, self-renewal, and differentiation [19–21].

In recent studies using of bioreactors, scaffolds, nanoparticles, and nanofibrils are diverse practical methods to improve expansion conditions of HSCs in vitro [22, 23].

Co-culturing of MSCs with HSCs is commonly used for improving their proliferation. Numerous parameters such as cytokines, extracellular vehicles (EVs), scaffolding, three-dimensional culture, the source of MSCs, and hypoxia affect the co-culturing of these two cell types [17, 22]. The culture conditions of these cells could be strengthened by manipulating these parameters. In this article, we reviewed the impact of those mentioned parameters on HSCs expansion. It might suggest practical strategies to provide more competent cells for therapeutic purposes.

Cellular secretions

Studies indicate that MSCs produce and secrete many chemokines, cytokines, growth factors, and extracellular matrix molecules. Microenvironmental signals are generated in MSC's niche, that can regulate various physiological and pathological processes including cell proliferation, differentiation, tissue development, regeneration and tumorigenesis [24, 25]. Cytokines and hematopoietic growth factors such as SDF, GM-CSF, Angs [26, 27], as well as substances secreted from MSCs, including micro vesicles (MVs) and exosomes, are valuable tools, utilized in vitro and in vivo researches in terms of basic sciences and disease treatment [28, 29]

Cytokines

MSCs play a critical role in the maintenance and differentiation of HSCs in the bone marrow niche by secreting cytokines and growth factors [30]. Several cytokines involve in the HSC differentiation and proliferation including IL-6, FLT3L, SCF, GCSF, and GM-CSF [6]. These cytokines and hematopoietic growth factors are utilized in different HSC expansion methods to provide a sufficient number of HSCs [6]. Using of MSC-feeder layer results in more cytokine production in the culture media. In these conditions, differentiation into the myeloid lineage is more likely to occur as an effect of cytokines [31]. In a study, co-culture of bone marrow MSCs with cord blood HSCs with a cytokine cocktail of FLT3L, SCF, basic fibroblast growth factor (BFGF), and leukemia inhibitory factor (LIF) led to further differentiation into the myeloid lineage. At the same time, a significant number of cells maintained their potential for lymphoid differentiation [32]. Cytokines widely use in the ex vivo expansion of HSCs because of their role in regulating the fate of HSCs. For this purpose, by adding exogenous cytokines such as SCF, FLT3L, and TPO in the culture media in co-culture of MSCs with HSCs, more clonogenic cells could be proliferated and expanded [33]. High levels of SDF in the MSC-HSC co-culture system reduce the secretion of inhibitor cytokines that negatively influences the self-renewal capacity of HSCs and progenitor cells [34]. Ahmadnejad et al. examined the culture of HSCs in different conditions, culture with MSCs feeder layer, culture with MSCs feeder layer plus cytokine cocktail and cytokine cocktail only. They concluded that using MSCs feeder layer with cytokine cocktail enhances cell proliferation [35]. Furthermore, cell proliferation with cytokines only results in the low survival of CD34 + cells [36], highlighting the importance of using MSCs in co-culture with HSCs.

In another study, the usage of MSCs as feeder layer and inhibition of transforming growth factor-beta receptor II (TGFβ-RII) increased the number of CD34⁺ cells, the total number of nucleated cells, and the number of colonies in vitro [37]. Therefore, using different cytokine cocktails in combination with MSC co-culture is one of the practical methods to optimize HSCs expansion conditions.

Microvesicles

Extracellular vesicles are cell-derived vesicles with a lipid bilayer membrane that ranges from 30 to 2000 nm based on their origin. Extracellular vesicles can be divided into three main classes: exosomes, micro vesicles, and apoptotic bodies, which differ in size and production mechanisms [38]. MVs establish intracellular communication and they also interact with target cells by controlling their biological functions [39]. They contain cytokines, signaling proteins, microRNAs (miRNAs), and mRNAs of origin cells. They could affect subjected cells by delivering these mentioned materials [40]. MSC-derived MVs play an essential role in maintaining intra-tissue homeostasis and could control cell responses to different stimuli especially in disease or injury [41]. A study by De Luca et al., reported that MSC-derived MVs affect the gene expression pattern of umbilical cord blood CD34⁺ cells and enhance their survival rate, inhibit their differentiation, and increase their migration to the bone marrow. miRNAs containing MVs interfere with the fate of umbilical cord blood CD34⁺ cells. MSC-derived MVs cooperate in the regeneration of the hematopoietic compartments, which can be a suitable therapeutic tool for the treatment of various hematologic and non-hematologic diseases [42]. Figure 1 represents the effect of MSC-secreted cytokines and MSC MVs on HSCs briefly (Fig. 1).

Scaffold and 3-dimensional culture

Studies indicate the importance of a three-dimensional culture system and scaffolds for simulating the cells physiological condition ex vivo [43, 44]. Despite its easy application, a Two-dimensional culture system, has some limitations such as low cell proliferation capacity. In two- dimensional culture, a monolayer of cells is grown, contrary to the three-dimensional physiological condition of the tissue in vivo. Improper cellular interaction and risk of hydrodynamic damages are other potential limitations of two-dimensional culture conditions. The purpose of three-dimensional culture is increasing the surface-to-volume ratio, the contact surface and cell communication [45, 46]. Scaffolds act as a physical supporter, which increases the proliferation rate and then the number of expanded HSCs [43]. Typically, a two-dimensional layer of MSCs is used in the MSC and HSC co-culture to support CD34⁺ cell proliferation. Two-dimensional culture does not adequately mimic bone marrow microenvironmental conditions and reduces the opportunities to cultivate engraftable HSCs with long-term culture ability [22, 47]. The three-dimensional culture of MSCs in a co-culture system not only increases their positive effects on the HSCs proliferation but also preserves HSCs self-renewal and stemness potentialities [48]. One of the strategies to strengthen MSCs and HSCs co-culture systems is the use of scaffolds to create a three-dimensional dynamic microenvironment [19]. Bioreactors, nanoparticles, nanofibrils, and scaffolds could apply to improve HSCs cultivation conditions [49–51]. For example, in one study, fibronectin-coated polycaprolactone scaffolds were used to expand HSCs. The results showed an increase in CD34⁺ cell number compared to the two-dimensional culture. Mousavi et al. isolated HSCs from the umbilical cord blood and cultured these cells in the presence of polycaprolactone nano scaffolds containing fibronectin. Cell enumeration results showed that the number of cells cultured with nano scaffold was much higher than the control group (culture under normal conditions without nano scaffold). Moreover, the expanded cells on the scaffolds had more clonogenicity [49, 50]. Futerga et al. reported that co-culture of MSC and HSCs in Poly dimethyl siloxane Microwells (PDMS) increased the number of CD34⁺ CD38⁻ cells, which was independent on MSCs but in response to PDMS properties [20]. In another study, MSC and HSC co-culture systems were established without using cytokines and other bioactive compounds in three-dimensional culture with bone-made –scaffolds. The result showed significant increase of HSCs in this condition compared to two-dimensional culture [21].

Hypoxia

As mentioned earlier, HSCs locate in the bone marrow in particular microenvironments known as the HSC niche [16]. Stem cell niche has low oxygen pressure [40]. In physiological conditions, stem cells distribute in different tissues with low oxygen pressure [18]. The culture of mouse and human HSCs at 20% oxygen pressure causes stem cell depletion, while culture under 1% oxygen condition causes better cell survival [52]. The mechanism of HSC response to hypoxia conditions is probably related to hypoxia-inducible factor 1 (HIF-1), which activates at low oxygen pressure [53]. This situation changes the metabolic pathway from oxidative to glycolytic one, reduces the production of reactive oxygen species (ROSs), and enhances the degradation of these molecules. ROSs inhibit HSC differentiation [54]. Hypoxia seems to protect cells from oxidative stress damage [55]. Zhao et al. co-cultured Wharton jelly-derived MSCs with HSCs under 1% oxygen pressure (hypoxia) or 20% (normoxia), without the addition of external cytokines. Their results revealed that at 1% oxygen pressure, the number of CD34⁺ Lin⁻ primary cells increased significantly [56]. 1% oxygen pressure reduces the differentiation and maintains the pluripotency of HSCs, while 3–5% oxygen pressure does not affect cell differentiation [57]. Mohammadali et al. found that mild hypoxia induces more rapid proliferation in HSCs and increases the number of expanded total nucleated cells. Moreover, expansion of human CD34⁺/ CD38⁻ cells under hypoxia conditions increases engraftment rate. Transplantation of hypoxia-expanded-HSCs leads to more reconstitution of hematopoietic tissue in mice in vivo [58].

Use of fetal-tissue-derived MSCs in co-culture with HSCs

Wharton jelly (WJ)-derived MSCs

WJ-MSCs as an easy-access source of MSCs have recently emphasized the attention of researchers due to their fewer ethical concerns. The population of WJ-MSCs multiplies faster than bone marrow-derived MSCs (BM-MSCs). These cells are more primitive in comparison with bone marrow MSCs and probably are also less immunogenic [59]. WJ-MSCs can be an ideal candidate for usage as feeder layer in a co-culture system due to their high proliferation capacity and rapid growth. WJ-MSCs express markers such as CD117 ([c-kit), and SCF receptors at a high level that helps to identify them in the co-culture with HSCs [60]. Osteopontin is highly expressed in WJ-MSCs [61]. On the other hand, WJ-MSCs can secrete hyaluronic acid [62]. Both of these molecules are significant constituents of HSC's niche. In particular, osteopontin is an essential regulator of HSC quiescence and proliferation. WJ-MSCs, like other MSCs, express CXCL12, which interacts with its receptor, CXCR4, to regulate HSC migration and homing. WJ-MSCs secrete most of the cytokines involved in the regulation of hematopoiesis. The concentration of hematopoietic growth factors such as G-CSF and GM-CSF secreted by these cells is higher than the factors secreted by MSCs isolated from other sources such as bone marrow [63, 64]. Then it seems that the application of WJ-MSCs as feeder layer might improve the expansion condition of HSCs.

Umbilical cord/ Umbilical cord blood-derived MSCs

BM-MSCs isolation is an invasive and painful procedure. But MSCs can isolate from embryo associated tissue, such as the umbilical cord (UC) or umbilical cord blood (UCB). UCB and UC are rich sources of MSCs and be considered for isolation of these valuable cells [65] UC-MSCs secrete more than 200 biologically active molecules, including cytokines, chemokines, and hematopoietic growth factors [66]. Therefore, UC used as an alternative source of MSC for clinical applications [67]. UC-MSCs are more initial/ primary than BM-MSCs. In addition, UC-MSCs proliferate faster than BM-MSCs. In the study of Wu et al., a sufficient number of UC-MSCs obtained for transplantation within eight days, which is shorter than the time required for BM-MSC harvesting [68]. UC-MSCs do not express HLA class II molecules on their surface. But they express B7-H1, a negative regulator of the immune system. Then they also prevent the reaction and activation of T lymphocytes and dendritic cells [69]. They also release indoleamine 2,3-dioxygenase and IL-12 suppress the immune system and increase graft survival [70]. These cells enhance the proliferation of regulatory T lymphocytes and are effective in treating graft versus host disease [71]. UC-MSCs and UCB-MSCs secrete SDF-1, Flt3, TPO, G-CSF, GM-CSF, IL6, SCF, and LIF, which play a vital role in supporting HSC growth. Gao et al. found that UC-MSCs promote HSC growth rate and elevate the expansion of megakaryocyte colony-forming units (CFU-Mk) even more efficiently than BM-MSCs [72]. The use of embryonic-tissue- derived-MSCs in the co-culture system with the HSCs can increase the efficiency of the HSCs expansion method and provide a sufficient number of the cells for transplantation. Examples of co-culture invigorating factors and their mechanisms have been summarized in Table 1.

Table 1

Use of scaffolds, hypoxia, and umbilical cord/ umbilical cord blood-derived MSCs to strengthen MSC and HSC co-culture system and their effect.
Factors		Mechanism	Effect	Reference
Scaffold	PCL scaffold coated with FN	↑ CXCR-4, VLA-4, VLA-5, LFA-1, HOXB-4, HOXA-9, BMI-1, Htert ↑CD13, CD14, CD33, CD34, CD45 ↓CD2, CD3, CD19	↑Total number cells ↑Proliferation ↑Adhesion ↑Homing ↑Self-renewal	[49, 50]
	Polydimethylsiloxane microwells in the PDMS	↑CD34, ↓CD38	↑Homing	[20]
	Biomimetic macroparous PEG hydrogels 3D scaffold	Mimicking the HSC niche	↑Stemness	[51]
Hypoxia	Hypoxia tension 1% O₂	↑ VEGF, ↓ IL-6, ↓IL-7, ↓ SCF, ↓TPO of WJ-MSCs Activation of NOTCH, Wnt/βCatenin, Hedgehog signaling pathway	↑Total number cells ↑Stemness ↑CFUs ↑CD34⁺Lin^_ Cell s	[56]
Hypoxia	MSC feeder + cytokines and 5% O₂ tension	↑CD34 ↑ CXCR4 gene ↑ CXCR4 mRNA	↑Total number cells ↑Homig ↑CFUs ↑Migration	[58]
Embryo-dependent-tissue-derived MSCs	UCB-derived MSCs	↑ CD34 ↑ TPO ↓ GM-CSF, SCF	↑Proliferation of CD34⁺cells, ↑CFU-GM, ↑CFU-E ↑CFU-GM	[72]
Embryo-dependent-tissue-derived MSCs	WJ-derived MSCs	↑CD34	↑HSC expansion	[64]

Use of chemicals to strengthen MSC and HSC co-culture system

In most cases, the ex vivo culture of HSCs requires the addition of cytokines. However, cytokines often lead to the accumulation of ROSs in the HSCs. Accumulation of ROSs causes excessive oxidation of nucleic acids, lipids, amino acids, and carbohydrates. It not only induces senescence and apoptosis in the CD34⁺ population but also leads to carcinogenesis and impaired hematopoiesis [73, 74]. Then providing some other HSC expansion stimulators might be effective in this regard.

Chemical and herbal molecules use to improve the co-culture conditions of HSCs and MSCs. Resveratrol is a natural polyphenol found in various plants and fruits, including peanuts, berries, and grapes [75]. Resveratrol has various functions such as reducing oxidative stress, influencing the cell cycle, and binding to receptors on the HSCs surface. Therefore, it considers as a culture medium additive, which increases HSCs proliferation and improves their cultivation conditions [76].

Tang et al., proved that the addition of 10 µM resveratrol can induce CD34⁺ cell proliferation ex vivo, maintaining primary HSCs (CD34⁺/ CD38⁻/ CD45R⁻/ CD49f⁻/ CD90⁺ cells), and inhibits their apoptosis [77]. 1 µM resveratrol reduces ROSs and nitric oxide levels in HSCs. It might more effective in the co-culture of HSCs with MSCs and protects HSCs against apoptosis [78]. Copper involves in gene expression regulation, protein function, and cell proliferation and differentiation. However, it catalyzes ROSs production. Then an excessive amount of intercellular copper induces harsh toxic conditions. Zaker et al. showed that the addition of TEPA, a linear polyamine copper chelator, to the culture medium of HSC and MSC co-culture; while reducing the amount of intercellular copper, induces more proliferation of CD34⁺ cells compared to other culture conditions. TEPA preserves HSCs storage pool in bone marrow by reducing oxidative stress [79]. Zhang et al. studied the use of some chemical components and small molecules to increase HSC proliferation. In this investigation, it was shown that various chemicals including SR1 (aryl hydrocarbon receptor antagonist), UM171 (pyrimidoindole derivatives), P18IN003 ,and P18IN011 (p18 inhibitory molecules in the cell cycle), CHIR99021 (type of glycogen synthase kinase 3β inhibitor), rapamycin (mTOR inhibitor), BIO (another kind of glycogen synthase kinase 3β inhibitor), NR-101 (a small non-peptidyl agonist molecule of c-MPL), 5azaD / TSA (5-aza-2-deoxycytidine / trichostatin A), and GAR (garcinol; a plant-derived histone acetyltransferase) could increase HSC self-renewal capacity [80]. Both VPA (Valproic acid) and DEAB (diethylaminobenzaldehyde) by preventing HSCs differentiation and zVADfmk / zLLYfmk (caspase and calpain inhibitors) and 5-HT (5hydroxytryptamine or serotonin) by inhibiting HSC apoptosis enhance HSCs proliferation [80]. Adding these materials to the culture medium of HSC and MSC co-culture might be effectively optimizing the HSC cultivation condition and expansion. Table 2 represents mechanisms and effects of using chemicals in MSC and HSC co–culture system that has reported in different studies.

Table 2

Using chemical and herbal molecules to strengthen MSC and HSC co–culture system and their effects.
Kind of Chemical/ Component	Mechanism	Effect	References
Resveratrol	↓ Intracellular ROS level; ↑Strengthening the scavenging of ROS ↓Percentages of apoptotic cells in cultures	↑ Expansion of CD34⁺ cells Preserved more primitive HSCs ↑Biological function	[76–78]
TEPA	↑ Cellular Copper content	↑ HSC Expansion	[79]
TEPA	↓ Oxidative stress	HSC storage pool preservation	[79]
SR1	↓ AhR	↑ Number of CD34⁺ cells	[81]
UM171	↑Genes encoding surface molecules [TMEM183A and PROCR]	↑ Human HSC self-renewal	[82]
P18IN003 P18IN011	↓ p18^INK4C	↑The frequency of primitive hematopoietic cells culture	[83]
CHIR99021 Rapamycin	↓ mTOR ↑Wnt/β-catenin and PTEN/PI3K/AKT/mTOR	↑ HSCs ↑ Cell cycle progression	[84, 85]
BIO	↓ GSK3β	↑ HSC self-renewal	[86]
NR-101	STAT5 activation	↑ Human HSC self-renewal	[80]
NR-101	HIF-1α stabilization	Regulate the oxygen level Response to hypoxia ↑ Expression of VEGF	[80]
5azaD TSA	↓ DNA methyltransferase	↑ HSC self-renewal	[87]
GAR	↓ HAT	↑ HSC self-renewal	[88]
VPA	↓ HDAC	↓ HSC differentiation	[89]
DEAB	↓ ALDH ↓ Expression of cEBPε and HoxB4	↓ HSC differentiation	[90]
5-HT	5-HTR activation ↓ Caspase-3 level	↓ HSC apoptosis	[91]
zVADfmk zLLYfmk	↓ Caspase activity ↑ Bcl-2 ↓ Caspase-3 Notch1 activation	↓ HSC apoptosis	[92, 93]

ROS: Reactive oxygen species. AhR: Aryl hydrocarbon receptor. TMEM183A: Transmembrane protein 183A. PROCR: Protein C receptor. mTOR: Mammalian target rapamycin. PTEN: Phosphatase and tensin homolog. PI3K: Phosphoinositide 3-kinase. GSK3β: Glycogen synthase kinase 3 beta. STAT5: Signal transducer and activator of transcription 5. HIF-1α: hypoxia inducible factor-1α. 5azaD: 5-Aza-2-Deoxycytidine. TSA: Trichostatin A.HAT: Histone acetyltransferase. HDAC: Histone deacetylase. DEAB: Diethylaminobenzaldehyde. ALDH: Aldehyde dehydrogenase. VPA: Valproic acid. GAR: Garnicol. 5-HT: 5-Hydroxytryptamine.

Non-adherent suspension cultivation

MSCs typically proliferate as monolayer adhere cells due to their intrinsic nature [5, 19]. Cell cultivation in the adherent phase could also cause some disadvantages and limitations. After a while, adherent culture in a monolayer phase led to decreased cell proliferation and expansion capacity [21]. The differentiation capacity and cellular plasticity of MSCs reduce during adhesive culture and multiple cell culture passages. The secretory capacity and the amount/ type of bioactive molecules secreted by MSCs often change and decrease gradually under adhesive conditions. On the other hand, the rate of cellular senescence increases in monolayer and adhesive culture [21, 24].

Non-adherent culture for colony/sphere formation is one of the practical methods to improve the differentiation and secretory capacities of MSCs [18, 25, 26]. In this case, cells proliferate in colony, and cell-cell contact increase during the culture period time. Then cell communication and signaling are performed effectively. The suspension culture of MSCs enhances their protective effects on HSCs by increasing paracrine properties [27].

MSC Non-adherent culture creates partial three-dimensional and dynamic culture condition and improves their proliferation, paracrine, differentiation abilities [26]. According to evidence, suspension cultivation of MSCs and MSC-spheres could be used as a simple method to enhance the efficacy of the co-culture system [18].

Nowadays, the use of MSCs and their derivatives have been considered in cell therapy fields to treat various diseases [57, 94]. The features of MSCs including, secretion of cytokines, migration/tropism to the injured tissues, immunomodulation, etc., have made them valuable tools for therapeutic approaches [95, 96]. HSC transplantation utilizes as a final treatment for autoimmune diseases, immunodeficiency, genetic disorder, hematopoietic tissue malignancies, and cancers [97]. Due to the therapeutic application of HSCs, the need for the effective expansion of these cells in vitro has increased [98]. Differentiation, proliferation, and stemness capacity of HSCs might reduce during their cultivation in vitro [47]. On the other hand, MSCs have supportive effects on HSCs and could use as a feeder layer [18]. Therefore, the co-culture of HSCs with MSCs is used to expand HSCs. By improving the co-culture system of these cells via examining the involved parameters, defining strategies to optimize it, and strengthening this culture method, we can harvest a sufficient number of competent cells for transplantation and other research uses. For this purpose, several studies have been conducted and different conditions of the co-culture have been investigated in terms of the presence of cytokines, use of scaffolds, adding various additives to the culture medium, modification on cultivation conditions, and oxygen pressure. Some of these conditions could be used in a combination manner [18]. These strategies were represented in Fig. 2.

Today, due to the therapeutic application of stem cells, especially HSCs, for the treatment of malignant and non-malignant diseases, the study of these cells has been considered in vitro. At the same time, in vivo studies have been designed to overcome the existing limitations. Indeed, the disadvantages and advantages of using these methods are not yet fully understood in the clinic. Therefore, more studies and researches should perform in this regard.

FLT3: Fetal liver tyrosine kinase-3.

FGF: Fibroblast Growth Factor.

HSC: Hematopoietic Stem Cell.

MV: Micro vesicle.

SCF: Stem Cell Factor.

LIF: Leukemia Inhibitory Factor.

TPO: Thrombopoietin.

PCL: Polycaprolactone. FN: Fibronectin.

PDMS: Polydimethyl siloxane.

PEG: Polyethilene glycol.

CFU: Colony-Forming Unit.

CFU-GM: Colony-forming-unit granulocyte/macrophage.

CFU-E: Colony-forming unit erythroid.

IL: Interleukine.

VLA: Very late antigen.

LFA: Lymphocyte function-associated antigen.

GM-CSF: Granulocyte-macrophage colony stimulating factor.

VEGF: Vascular endothelial growth factor.

UCB: Umbilical cord blood.

MSCs: Mesenchymal stem cell.

WJ: Wharton jelly.

HSC: Hematopoietic stem cell.

ROS: Reactive oxygen species.

TEPA: Tetraethylenepentamine.

AhR: Aryl hydrocarbon receptor.

TMEM183A: Transmembrane protein 183A.

PROCR: Protein C receptor.

mTOR: Mammalian target rapamycin.

PTEN: Phosphatase and tensin homolog.

PI3K: Phosphoinositide 3-kinase.

GSK3β: Glycogen synthase kinase 3 beta.

STAT5: Signal transducer and activator of transcription 5.

HIF-1α: hypoxia inducible factor-1α.

5azaD: 5-Aza-2-Deoxycytidine.

TSA: Trichostatin A.

GAR: Garnicol.

HAT: Histone acetyltransferase.

VPA: Valproic acid.

HDAC: Histone deacetylase.

DEAB: Diethylaminobenzaldehyde.

ALDH: Aldehyde dehydrogenase.

5-HT: 5-Hydroxytryptamine.

Funding:

This work was founded by the Hamadan University of Medical Sciences [grant No. 140004012854].

Competing interests:

All authors contributed in this review clearly state that there is no conflict of interest either in authorship or financial benefits.

Authors contributions:

Mandana Shirdarreh has written the first draft of the manuscript. Mohammad Pouya Samieeand Armita Safarihave participated to write the first manuscript and figure preparation. Fatemeh Amiri has reviewed the text. All authors commented on previous versions of the manuscript. All authors reviewed the manuscript.

Ethics approval:

This is a review article. The research ethic committee of Hamadan University of Medical Sciences has confirmed that no ethical approval is required.

Consent to participate:

Not applicable.

Consent to publish:

All authors reviewed the manuscript and consent to publish

Acknowledgment

This work was founded by the Hamadan University of Medical Sciences, Code: IR.UMSHA.REC.1400.183, [grant number: 140004012854]. Special thanks to Delaram Yaghoubzadeh.

Molaei S, Amiri F, Salimi R et al (2022) Therapeutic effects of mesenchymal stem cells-conditioned medium derived from suspension cultivation or silymarin on liver failure mice. Mol Biol Rep
Kokkaliaris KD, Scadden DT (2020) Cell interactions in the bone marrow microenvironment affecting myeloid malignancies. Blood Adv 415:3795–3803
Nakahara F, Borger DK, Wei Q et al (2019) Engineering a haematopoietic stem cell niche by revitalizing mesenchymal stromal cells. Nat Cell Biol 215:560–567
Mushahary D, Spittler A, Kasper C et al (2018) Isolation, cultivation, and characterization of human mesenchymal stem cells. Cytometry Part A931:19–31
Schmelzer E, McKeel DT, Gerlach JC (2019) Characterization of human mesenchymal stem cells from different tissues and their membrane encasement for prospective transplantation therapies. Biomed Res Int
Pittenger MF, Mackay AM, Beck SC et al (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284:143–147
Al-Anazi K (2020) The Future Role of Mesenchymal Stem Cells in Tissue Repair and Medical Therapeutics: Realities and Expectations. J Reg Med Biol Res 12:1–5
Noort W, Scherjon S, Kleijburg-Van Der Keur Cet al et al (2003) Mesenchymal stem cells in human second-trimester bone marrow, liver, lung, and spleen exhibit a similar immunophenotype but a heterogeneous multilineage differentiation potential. Haematologica 888:845–852
Meng X, Ichim TE, Zhong J et al (2007) Endometrial regenerative cells: a novel stem cell population. J Transl Med 51:1–10
Goodarzi A, Valikhani M, Amiri F et al (2022) The mechanisms of mutual relationship between malignant hematologic cells and mesenchymal stem cells: Does it contradict the nursing role of mesenchymal stem cells? Cell Commun Signal 20:21–34
Saleh M, Shams Asanjan K, Movassaghpour Akbari A et al (2015) The Effect of Mesenchymal Stem Cells on Hematopoetic Stem Cells Differentiation. Sci J Iran Blood Transfus Organ 123:292–302
Amiri F, Molaei S, Bahadori M et al (2016) Autophagy-modulated human bone marrow-derived mesenchymal stem cells accelerate liver restoration in mouse models of acute liver failure. Iran Biomed J 203:135
Orkin SH, Zon LI (2018) SnapShot: hematopoiesis Cell1324:712. e1-. e2
Mangel M, Bonsall MB [2013]. Stem cell biology is population biology: differentiation of hematopoietic multipotent progenitors to common lymphoid and myeloid progenitors.Theor Biol Med Model, 17; 10:5
Wilson A, Trumpp A (2006) Bone-marrow haematopoietic-stem-cell niches. Nat Rev Immunol 62:93–106
Gomariz A, Isringhausen S, Helbling PM et al (2020) Imaging and spatial analysis of hematopoietic stem cell niches. Ann N Y Acad Sci 14661:5–16
Hurwitz SN, Jung SK, Kurre P (2020) Hematopoietic stem and progenitor cell signaling in the niche. Leukemia 34:3136–3148
Amiri F, Kiani AA, Bahadori M et al (2021) Co-culture of mesenchymal stem cell spheres with hematopoietic stem cells under hypoxia: a cost-effective method to maintain self-renewal and homing marker expression. Mol Biol Rep
Zhang P, Zhang C, Li J et al (2019) The physical microenvironment of hematopoietic stem cells and its emerging roles in engineering applications. Stem Cell Res Ther 10:1–13
Futrega K, Atkinson K, Lott WB et al (2017) Spheroid coculture of hematopoietic stem/progenitor cells and monolayer expanded mesenchymal stem/stromal cells in polydimethylsiloxane microwells modestly improves in vitro hematopoietic stem/progenitor cell expansion. Tissue Eng Part C Methods 23:200–218
Huang X, Zhu B, Wang X et al (2016) Three-dimensional co-culture of mesenchymal stromal cells and differentiated osteoblasts on human bio-derived bone scaffolds supports active multi-lineage hematopoiesis in vitro: Functional implication of the biomimetic HSC niche. Int J Mol Med 38:1141–1151
Khong D, Li M, Singleton A et al (2018) Stromalized microreactor supports murine hematopoietic progenitor enrichment. Biomed Microdevices 20:1–9
Mayle A, Luo M, Jeong M et al (2013) Flow cytometry analysis of murine hematopoietic stem cells. Cytometry Part A 83:27–37
Abbasi-Malati Z, Roushandeh AM, Kuwahara Y et al (2018) Mesenchymal stem cells on horizon: a new arsenal of therapeutic agents. Stem Cell Rev Rep 14::484–499
Soltani F, Amiri F, Mohammadipour M et al (2016) Cell survival evaluation of mesenchaymal stem cells cultivated in the presense of secretome of HIF-1α/Nrf2-engineered-MSC. Sci J Iran Blood Transfus Organ 124:318–330
Aqmasheh S (2017) Effects of mesenchymal stem cell derivatives on hematopoiesis and hematopoietic stem cells. Adv pharm bull 7:165
Qiu G, Zheng G, Ge M et al (2018) Mesenchymal stem cell-derived extracellular vesicles affect disease outcomes via transfer of microRNAs. Stem Cell Res Ther 9:1–9
Sagaradze G, Grigorieva O, Nimiritsky P et al (2019) Conditioned medium from human mesenchymal stromal cells: towards the clinical translation. Int J Mol Sci 20:1656
Robinson SN, Ng J, Niu T et al (2006) Superior ex vivo cord blood expansion following co-culture with bone marrow-derived mesenchymal stem cells. Bone Marrow Transplant 37:359–366
Zhang Y, Shen B, Guan X (2019) Safety and efficacy of ex vivo expanded CD34 + stem cells in murine and primate models. Stem Cell Res Ther 10:1–16
Lazar-Karsten P, Dorn I, Meyer G et al (2011) The influence of extracellular matrix proteins and mesenchymal stem cells on erythropoietic cell maturation. Vox Sang 1011:65–76
Andrade PZ, dos Santos F, Almeida-Porada G et al (2010) Systematic delineation of optimal cytokine concentrations to expand hematopoietic stem/progenitor cells in co-culture with mesenchymal stem cells. Mol Biosyst 6:1207–1215
Schuster JA, Stupnikov MR, Ma G et al (2012) Expansion of hematopoietic stem cells for transplantation: current perspectives. Exp Hematol Oncol 1:1–6
Ajami M, Soleimani M, Abroun S et al (2019) Comparison of cord blood CD34 + stem cell expansion in coculture with mesenchymal stem cells overexpressing SDF-1 and soluble/membrane isoforms of SCF. J Cell Biochem 120:15297–15309
Ahmadnejad M, Amirizadeh N, Mehrasa R et al (2017) Elevated expression of DNMT1 is associated with increased expansion and proliferation of hematopoietic stem cells co-cultured with human MSCs. Blood Res 52:25
Noort WA, Kruisselbrink AB, in't Anker PS et al (2002) Mesenchymal stem cells promote engraftment of human umbilical cord blood–derived CD34 + cells in NOD/SCID mice. Exp Hemato 30:870878
Akhkand SS, Amirizadeh N, Nikougoftar M et al (2016) Evaluation of umbilical cord blood CD34 + hematopoietic stem cells expansion with inhibition of TGF-β receptorII in co-culture with bone marrow mesenchymal stromal cells. Tissue Cell 484:305–311
Minciacchi VR, You S, Spinelli C, Morley S et al (2015) Large oncosomes contain distinct protein cargo and represent a separate functional class of tumor-derived extracellular vesicles. Oncotarget 6:11327
Robbins PD, Morelli AE (2014) Regulation of immune responses by extracellular vesicles. Nat Rev Immunol 14:195–208
Collino F, Deregibus MC, Bruno S (2010) Micro vesicles derived from adult human bone marrow and tissue specific mesenchymal stem cells shuttle selected pattern of miRNAs.PloS one5:e11803
Asgarpour K, Shojaei Z, Amiri F et al (2020) Exosomal microRNAs derived from mesenchymal stem cells: cell-to-cell messages. Cell Commun Signal 18:1–16
De Luca L, Trino S, Laurenzana I (2016) MiRNAs and piRNAs from bone marrow mesenchymal stem cell extracellular vesicles induce cell survival and inhibit cell differentiation of cord blood hematopoietic stem cells: a new insight in transplantation. Oncotarget 7:6676
Jing D, Fonseca AV, Alakel N et al (2010) Hematopoietic stem cells in coculture with mesenchymal stromal cells-modeling the niche compartments in vitro. Haematologica 95:542–550
Miyoshi H, Shimizu Y, Yasui Y et al (2020) Expansion of mouse primitive hematopoietic cells in three-dimensional cultures on chemically fixed stromal cell layers. Cytotechnology 72:741–750
Jozaki T, Aoki K, Mizumoto H et al (2010) In vitro reconstruction of a three-dimensional mouse hematopoietic microenvironment in the pore of polyurethane foam. Cytotechnology 62:531–537
Aggarwal R, Lu J, Pompili VJ et al (2012) Hematopoietic stem cells: transcriptional regulation, ex vivo expansion and clinical application. Curr Mol Med 12:3449
Robinson SN, Simmons PJ, Yang H et al (2011) Mesenchymal stem cells in ex vivo cord blood expansion. Best Pract Res Clin Haematol 24:83–92
De Lima M, Mcniece I, Robinson SN et al (2012) Cord-blood engraftment with ex vivo mesenchymal-cell coculture. N Engl J Med 367:23052315
Mousavi SH, Abroun S, Soleimani M et al (2018) 3-Dimensional nano-fibre scaffold for ex vivo expansion of cord blood haematopoietic stem cells. Artif Cells Nanomed Biotechnol 46:740–748
Mousavi SH, Abroun S, Soleimani M et al (2015) Expansion of human cord blood hematopoietic stem/progenitor cells in three-dimensional Nano scaffold coated with Fibronectin. Int J Hematol Oncol Stem Cell Res 9:72
Raic A, Rödling L, Kalbacher H et al (2014) Biomimetic macroporous PEG hydrogels as 3D scaffolds for the multiplication of human hematopoietic stem and progenitor cells. Biomaterials 35:929–940
Hermitte F, de la Brunet P, Belloc F et al (2006) Very low O2 concentration [0.1%] favors G0 return of dividing CD34 + cells. Stem Cells 24:65–73
Zhang CC, Sadek HA (2014) Hypoxia and metabolic properties of hematopoietic stem cells. Anti-oxide Redox Signal 20:1891–1901
Suda T, Takubo K, Semenza GL (2011) Metabolic regulation of hematopoietic stem cells in the hypoxic niche. Cell Stem Cell 9:298–310
Jang YY, Sharkis SJ (2007) A low level of reactive oxygen species selects for primitive hematopoietic stem cells that may reside in the low-oxygenic niche. Blood 110:3056–3063
Zhao D, Liu L, Chen Q et al (2018) Hypoxia with Wharton’s jelly mesenchymal stem cell coculture maintains stemness of umbilical cord blood-derived CD34 + cells. Stem Cell Res Ther 9:1–11
Ezashi T, Das P, Roberts RM (2005) Low O2 tensions and the prevention of differentiation of hES cells. Proc Natl Acad Sci 102:4783–4788
Mohammadali F, Abroun S, Atashi A (2018) Mild hypoxia and human bone marrow mesenchymal stem cells synergistically enhance expansion and homing capacity of human cord blood CD34 + stem cells. Iran J Basic Med Sci 21::709
Amiri F, Halabian R, Dehgan Harati M et al (2015) Positive selection of Wharton's jelly-derived CD105 + cells by MACS technique and their subsequent cultivation under suspension culture condition: A simple, versatile culturing method to enhance the multipotentiality of mesenchymal stem cells. Hematol 20:208–216
Anzalone R, Iacono ML, Corrao S et al (2010) New emerging potentials for human Wharton’s jelly mesenchymal stem cells: immunological features and hepatocyte-like differentiative capacity. Stem Cells Dev 194:423–438
Nilsson SK, Johnston HM, Whitty GA et al (2005) Osteo pontin, a key component of the hematopoietic stem cell niche and regulator of primitive hematopoietic progenitor cells. Blood 106:1232–1239
Raio L, Cromi A, Ghezzi F et al (2005) Hyaluronan content of Wharton's jelly in healthy and Down syndrome fetuses. Matrix Biol 24:166–174
Li Q, Zhao D, Chen Q et al (2019) Wharton’s jelly mesenchymal stem cell-based or umbilical vein endothelial cell-based serum-free coculture with cytokines supports the ex vivo expansion/maintenance of cord blood hematopoietic stem/progenitor cells. Stem Cell Res Ther 101:1–9
Lo Iacono M, Russo E, Anzalone R et al (2018) Wharton’s Jelly Mesenchymal Stromal Cells Support the Expansion of Cord Blood–derived CD34 + Cells Mimicking a Hematopoietic Niche in a Direct Cell–cell Contact Culture System. SAGE Publications Sage CA, Los Angeles, CA
Raffo D, Maglioco A, Fernandez Sasso D (2021) A protocol for umbilical cord tissue cryopreservation as a source of mesenchymal stem cells. Mol Biol Rep 48(2):1559–1565
Romanov YA, Volgina NE, Balashova EE et al (2017) Human umbilical cord mesenchymal stromal cells support viability of umbilical cord blood hematopoietic stem cells but not the “stemness” of their progeny in co-culture. Bull Exp Biol Med 163:523–527
Dai Y, Cui X, Zhang G et al (2022) Development of a novel feeding regime for large scale production of human umbilical cord mesenchymal stem/stromal cells. Cytotechnology 74:351–369
Wu KH, Tsai C, Wu HP et al (2013) Human application of ex vivo expanded umbilical cord-derived mesenchymal stem cells: enhance hematopoiesis after cord blood transplantation. Cell Transpl 22:2041–2051
Xu M, Zhang B, Liu Y et al (2014) The immunologic and hematopoietic profiles of mesenchymal stem cells derived from different sections of human umbilical cord. Acta Biochim Biophys Sin 46:1056–1065
Tipnis S, Viswanathan C (2010) Umbilical cord matrix derived mesenchymal stem cells can change the cord blood transplant scenario. Int J Stem Cells 3::103
Yang S, Wei Y, Sun R et al (2020) Umbilical cord blood-derived mesenchymal stromal cells promote myeloid-derived suppressor cell proliferation by secreting HLA-G to reduce acute graft-versus-host disease after hematopoietic stem cell transplantation. Cito therapy 22:718–733
Gao L, Chen X, ZhAng X et al (2006) Human umbilical cord blood-derived stromal cell, a new resource of feeder layer to expand human umbilical cord blood CD34 + cells in vitro. Blood Cells Mol Dis 36::322–328
Chen Y, Liang Y, Luo X et al (2020) Oxidative resistance of leukemic stem cells and oxidative damage to hematopoietic stem cells under pro-oxidative therapy. Cell Death Dis 11:1–12
Hu L, Zhang Y, Miao W et al (2019) Reactive oxygen species and Nrf2: functional and transcriptional regulators of hematopoiesis. Oxid Med Cell Longev 2019: 2019
Baur JA, Sinclair DA (2006) Therapeutic potential of resveratrol: the in vivo evidence. Nat Rev Drug Discov 5:493–506
Heinz N, Ehrnström B, Schambach A et al (2015) Comparison of different cytokine conditions reveals resveratrol as a new molecule for ex vivo cultivation of cord blood-derived hematopoietic stem cells. Stem Cells Transl Med 4:1064–1072
Tang C, Zhang W, Cai H et al (2019) Resveratrol improves ex vivo expansion of CB-CD34 + cells via downregulating intracellular reactive oxygen species level. J Cell Biochem 120:7778–7787
Vaidya A, Kale V, Poonawala M et al (2018) Mesenchymal stromal cells enhance the hematopoietic stem cell-supportive activity of resveratrol. Regen Med 13:409–425
Zaker F, Nasiri N, Oodi A et al (2013) Evaluation of umbilical cord blood CD34 + hematopoietic stem cell expansion in co-culture with bone marrow mesenchymal stem cells in the presence of TEPA. Hematology 18:39–45
Zhang Y, Gao Y (2016) Novel chemical attempts at ex vivo hematopoietic stem cell expansion. Int J Hematol 103:519–529
Singh KP, Casado FL, Opanashuk LA et al (2009) The aryl hydrocarbon receptor has a normal function in the regulation of hematopoietic and other stem/progenitor cell populations. Biochem Pharmacol 77:577–587
Fares I, Chagraoui J, Gareau Y et al (2014) Cord blood expansion Pyrimidoindole derivatives are agonists of human hematopoietic stem cell self-renewal. Science 345:1509–1512
Gao Y, Yang P, Shen H et al (2015) Small molecule inhibitors targeting INK4 protein p18[INK4C] enhance ex vivo expansion of haematopoietic stem cells. Nat Commun 6::6328
Chen C, Liu Y, Liu R et al (2008) TSC-mTOR maintains quiescence and function of hematopoietic stem cells by repressing mitochondrial biogenesis and reactive oxygen species. J Exp Med 205::2397–2408
Ying QL, Wray J, Nichols J et al (2008) The ground state of embryonic stem cell self-renewal. Nature 453:519–523
Ko KH, Holmes T, Palladinetti P et al (2011) GSK-3β inhibition promotes engraftment of ex vivo-expanded hematopoietic stem cells and modulates gene expression. Stem Cells 29:108–118
Araki H, Yoshinaga K, Boccuni P et al (2007) Chromatin-modifying agents permit human hematopoietic stem cells to undergo multiple cell divisions while retaining their repopulating potential. Blood 109:3570–3578
Nishino T, Wang C, Mochizuki-Kashio M et al (2011) Ex vivo expansion of human hematopoietic stem cells by garcinol, a potent inhibitor of histone acetyltransferase. PLoS ONE 6:e24298
De Felice L, Tataelli C, Mascolo MG et al (2005) Histone deacetylase inhibitor valproic acid enhances the cytokine-induced expansion of human hematopoietic stem cells. Cancer Res 65:1505–1513
Chute JP, Muramoto GG, Whitesides J et al (2006) Inhibition of aldehyde dehydrogenase and retinoid signaling induces the expansion of human hematopoietic stem cells. Proc Natl Acad Sci U S A 103:1707–1712
Yang M, Li K, Ng PC et al (2007) Promoting effects of serotonin on hematopoiesis: ex vivo expansion of cord blood CD34 + stem/progenitor cells, proliferation of bone marrow stromal cells, and antiapoptotic. Stem Cells 25:1800–1806
Kale VMS, Limaye VP LS (2010) Expansion of cord blood CD34 cells in presence of zVADfmk and zLLYfmk improved their in vitro functionality and in vivo engraftment in NOD/SCID mouse. PLoS ONE 58:e12221
Ogilvy S, Metcalf D, Print CG et al (1999) Constitutive Bcl-2 expression throughout the hematopoietic compartment affects multiple lineages and enhances progenitor cell survival. Proc Natl Acad Sci U S A 96:14943–14948
Driscoll J, Patel T (2019) The mesenchymal stem cell secretome as a cellular regenerative therapy for liver disease. J gastroenterol 54:763–773
Jiang H, Wang H, Liu T (2018) Co-cultured the MSCs and cardiomyocytes can promote the growth of cardiomyocytes. Cytotechnology 70:793–806
Bazinet A, Popradi G (2019) A general practitioner’s guide to hematopoietic stem-cell transplantation. Curr Oncol 26:187
Takagaki S, Yamashita R, Hashimoto N et al (2019) Galactosyl carbohydrate residues on hematopoietic stem/progenitor cells are essential for homing and engraftment to the bone marrow. Sci Rep 9:1–11
Feng Q, Chai C, Jiang XS et al (2006) Expansion of engrafting human hematopoietic stem/progenitor cells in three-dimensional scaffolds with surface immobilized fibronectin. J Biomed Mater Res A 78:781–791

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Influential factors for optimizing and strengthening mesenchymal stem cells and hematopoietic stem cells co-culture

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Journal Publication

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Abstract

Figures

Background

Cellular secretions

Cytokines

Microvesicles

Scaffold and 3-dimensional culture

Hypoxia

Use of fetal-tissue-derived MSCs in co-culture with HSCs

Wharton jelly (WJ)-derived MSCs

Umbilical cord/ Umbilical cord blood-derived MSCs

Use of chemicals to strengthen MSC and HSC co-culture system

Non-adherent suspension cultivation

Discussion And Conclusion

Abbreviations

Declarations

References

Status:

Journal Publication

Version 1