An overview and synopsis of techniques for directing stem cell differentiation in vitro

Heng, Boon Chin; Cao, Tong; Haider, Husnain Kh; Wang, Dennis Zheng Ming; Sim, Eugene K.W.; Ng, Soon-Chye

doi:10.1007/s00441-003-0847-5

Cited by 61 publications

(38 citation statements)

References 155 publications

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“…Instead, we have used 2-deoxy-5-azacytidin, a demethylating agent that can be employed to prime cells before transplantation in vivo (Zhang et al 2007;Burlacu 2006) and that is thus possibly more suitable for clinical use. Such a priming of stem cells in vitro before transplantation might be important, since theoretically this would prevent the spontaneous differentiation of stem cells into undesired lineages (Heng et al 2004).…”

Section: Discussionmentioning

confidence: 99%

Differentiation of human adipose-derived stem cells towards cardiomyocytes is facilitated by laminin

et al. 2008

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Adipose-derived stem cells (ASCs) are promising candidates for therapy in myocardial infarction (MI). However, the frequency of human ASCs that differentiate towards cardiomyocytes is low. We hypothesized that adherence to extracellular matrix molecules that are upregulated after MI might increase human stem cell differentiation towards cardiomyocytes. We analysed putative ASC differentiation on fibronectin-coated, laminincoated and uncoated culture plates. Expression of cardiac markers in cells was analysed 1, 3 and 5 weeks after stimulation with 5-aza-2-deoxycytidine. After 1 week, mRNA expression of myosin light chain-2α (MLC-2α), an early marker in cardiomyocyte development, was increased significantly in treated cells, independent of coating. At 5 weeks, however, mRNA expression of the late cardiomyocyte development marker SERCA2α was only significantly increased in 5-aza-2-deoxycytidine-treated cells cultured on laminin. Significantly higher numbers of cells were immunopositive for MLC-2α in cultures of treated cells grown on laminin-coated wells, when compared with cultures of treated cells grown on uncoated wells, both at 1 week and at 5 weeks. Furthermore, after 3 weeks, significantly more α-actinin-and desmin-positive cells were detected after treatment with 5-aza-2-deoxycytidine, but only in uncoated wells. After 5 weeks, however, the number of desmin-positive cells was only significantly increased after treatment of cells with 5-aza-2-deoxycytidine and culture on laminin (61% positive cells). Thus, we have found that a high percentage of human ASCs can be differentiated towards cardiomyocytes; this effect can be improved by laminin, especially during late differentiation.

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Section: Discussionmentioning

confidence: 99%

Differentiation of human adipose-derived stem cells towards cardiomyocytes is facilitated by laminin

et al. 2008

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“…Morphology of the cells by phase-contrast microscopy (c) and Oil red O staining (d) after three cycles of adipogenesis induction (total 18 days) showing the increase in size of the lipid vacuoles, and the typical morphology of adipocytes. Bar 100 μm The techniques used for the isolation and differentiation MSCs were a combination of those previously used in other species together with minor modifications aimed at achieving a better expression of lineage markers (Ringe et al 2002;Martin et al 2002;Heng et al 2004).…”

Section: Discussionmentioning

confidence: 99%

Isolation and multilineage differentiation of bovine bone marrow mesenchymal stem cells

Bosnakovski¹,

et al. 2004

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The bone marrow harbors a population of mesenchymal stem cells (MSCs) that possess the potential to differentiate into bone, cartilage, and fat, and along other tissue pathways. To date, MSCs from various species have been studied. Despite the bovine experimental model being widely used in experiments in vivo and in vitro, only a limited amount of information regarding bovine MSCs is available. The aim of this study was to isolate and induce the multilineage mesenchymal differentiation of bovine MSCs, thereby initiating further research on these cells. Bovine MSCs were isolated from eight calves, and osteogenic, chondrogenic, and adipogenic differentiation was induced by using a combination of previously reported protocols for other species. The level of differentiation was evaluated by histological examination and by analyzing the expression of tissue-specific genes by a quantitative "real time" reverse transcription/polymerase chain reaction technique. Following osteoinduction, the isolated fibroblast-like cells transformed into cuboidal cells and formed alkaline-phosphatase-positive colonies; during differentiation, these colonies transformed into mineralized nodules. In addition, osteogenesis was followed by osteocalcin and collagen type I mRNA expression. Chondrogenesis was confirmed by the demonstration of collagen type II, aggrecan, and sox9 mRNA expression in the cells stimulated by transforming growth factor β1 in monolayer culture. After being cultured in an adipogenesis-inducing medium, the MSCs responded by the accumulation of lipid vacuoles and the expression of adipocyte-specific genes. We have therefore demonstrated that cells harvested from bovine bone marrow are capable of in vitro extensive multiplication and multilineage differentiation, making them a relevant and invaluable model in the field of stem cell research.

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“…Extracellular signaling involves the interaction of cell surface receptors with soluble cytokines and growth factors, with the extracellular matrix (ECM), such as collagen and proteoglycans, or with the surface proteins of the neighboring cells (Heng et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

Chondrogenic differentiation of bovine bone marrow mesenchymal stem cells (MSCs) in different hydrogels: Influence of collagen type II extracellular matrix on MSC chondrogenesis

Bosnakovski

Mizuno

Kim

et al. 2006

Biotech & Bioengineering

438

322

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Bone marrow mesenchymal stem cells (MSCs) are candidate cells for cartilage tissue engineering. This is due to their ability to undergo chondrogenic differentiation after extensive expansion in vitro and stimulation with various biomaterials in three-dimensional (3-D) systems. Collagen type II is one of the major components of the hyaline cartilage and plays a key role in maintaining chondrocyte function. This study aimed at analyzing the MSC chondrogenic response during culture in different types of extracellular matrix (ECM) with a focus on the influence of collagen type II on MSC chondrogenesis. Bovine MSCs were cultured in monolayer as well as in alginate and collagen type I and II hydrogels, in both serum free medium and medium supplemented with transforming growth factor (TGF) b1. Chondrogenic differentiation was detected after 3 days of culture in 3-D hydrogels, by examining the presence of glycosaminoglycan and newly synthesized collagen type II in the ECM. Differentiation was most prominent in cells cultured in collagen type II hydrogel, and it increased in a timedependent manner. The expression levels of the of chondrocyte specific genes: sox9, collagen type II, aggrecan, and COMP were measured by quantitative ''Real Time'' RT-PCR, and genes distribution in the hydrogel beads were localized by in situ hybridization. All genes were upregulated by the presence of collagen, particularly type II, in the ECM. Additionally, the chondrogenic influence of TGF b1 on MSCs cultured in collagenincorporated ECM was analyzed. TGF b1 and dexamethasone treatment in the presence of collagen type II provided more favorable conditions for expression of the chondrogenic phenotype. In this study, we demonstrated that collagen type II alone has the potential to induce and maintain MSC chondrogenesis, and prior interaction with TGF b1 to enhance the differentiation.

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An overview and synopsis of techniques for directing stem cell differentiation in vitro

Cited by 61 publications

References 155 publications

Differentiation of human adipose-derived stem cells towards cardiomyocytes is facilitated by laminin

Differentiation of human adipose-derived stem cells towards cardiomyocytes is facilitated by laminin

Isolation and multilineage differentiation of bovine bone marrow mesenchymal stem cells

Chondrogenic differentiation of bovine bone marrow mesenchymal stem cells (MSCs) in different hydrogels: Influence of collagen type II extracellular matrix on MSC chondrogenesis

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