Chondrogenic Differentiation of Cultured Human Mesenchymal Stem Cells from Marrow

Mackay, Alastair M.; Beck, Steve; Murphy, James M.; Barry, Frank; Chichester, C. O.; Pittenger, Mark F.

doi:10.1089/ten.1998.4.415

Cited by 1,190 publications

(983 citation statements)

References 49 publications

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“…The data demonstrated that sustained expression of TGF-β1 is more effective in inducing chondrogenic differentiation of human MSCs than IGF-1. Previous studies using TGF-β1 to induce chondrogenesis in MSC was done by addition of TGF-β1 protein [8,21,24]. The optimal concentration of TGF-β1 used in the protein form to induce chondrogenesis in MSCs was 10 ng/mL [21,24].…”

Section: Discussionmentioning

confidence: 99%

“…Previous studies using TGF-β1 to induce chondrogenesis in MSC was done by addition of TGF-β1 protein [8,21,24]. The optimal concentration of TGF-β1 used in the protein form to induce chondrogenesis in MSCs was 10 ng/mL [21,24]. Interestingly, the results of ELISA showed that chondrogenic differentiation was more evident in pellets made from cells transduced with AdTGF-β1 at 50 MOI.…”

Section: Discussionmentioning

confidence: 99%

“…The nontransduced cells were treated similarly. The FBScontaining medium was then replaced with defined medium, consisting of DMEM supplemented with ITS+ premix (BD Sciences, San Diego, CA,USA)as described previously [21,24].The premix contains 6.25 μg/mL insulin, 6.25 μg/mL transferrin, 6.25 μg/mL selenous acid, 5.35 μg/mL linoleic acid, and 1.25 μg/mL bovine serum albumin (BSA). Medium was also supplemented with 1 mM pyruvate, 37.5 μg/mL of ascorbate-2-phosphate, and 10 -7 M of dexamethasone.…”

Section: Pellet Culturesmentioning

confidence: 99%

“…Numerous studies have demonstrated that bone marrow-derived mesenchymal stem cells (MSCs) can give rise to chondrocytes under appropriate conditions [21][22][23]. Members of the TGF-β1 (transforming growth factor 1) super-family have been shown to play a role in the induction of chondrogenic differentiation of MSCs in vitro [21,24,25].…”

Section: Introductionmentioning

confidence: 99%

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Adenoviral-mediated transfer of TGF-β1 but not IGF-1 induces chondrogenic differentiation of human mesenchymal stem cells in pellet cultures

Kawamura¹,

Chu²,

Sobajima³

et al. 2005

Experimental Hematology

102

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Objective. The objective of the present study was to investigate the potential of application of growth factor genes to induce chondrogenic differentiation of human-derived mesenchymal stem cells (MSCs). The growth factor genes evaluated in the present study were transforming growth factor 1 (TGF-β1) and insulin-like growth factor 1 (IGF-1).Methods. Human MSCs were transduced with the adenoviral vectors carrying either TGF-β1 or IGF-1 (AdTGF-β1 and AdIGF-1 respectively) or a combination of both growth factor genes at different multiplicities of infection (MOI) and were then made into pellets. Pellets were also made from nontransduced cells and maintained in culture medium supplemented with 10 ng/mL of TGF-β1. At specified time points, histological analysis, cartilage matrix gene expression, and immunofluorescence were performed to determine the extent of chondrogenic differentiation.Results. MSCs transduced with the AdTGF-β1 demonstrated robust chondrogenic differentiation, while those made from AdIGF-1 did not. AdTGF-β1 pellets demonstrated aggrecan gene expression as early as day 3 of pellet culture, while type II collagen gene expression was detected by day 10 of culture. The AdIGF-1, alone or in combination with TGF-β1 pellets, did not show any type II collagen gene expression at any time point. By immunofluoresecence, type X collagen was distributed throughout the matrix in TGF-β1 protein pellets while the growth factor gene pellets displayed scant staining. Conclusion.The results suggest that sustained administration of TGF-β1 may be more effective in suppressing terminal differentiation than intermittent dosing and thus effective for cartilage repair.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Pellet Culturesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Adenoviral-mediated transfer of TGF-β1 but not IGF-1 induces chondrogenic differentiation of human mesenchymal stem cells in pellet cultures

Kawamura¹,

Chu²,

Sobajima³

et al. 2005

Experimental Hematology

102

View full text Add to dashboard Cite

show abstract

“…Bone marrow mesenchymal stem cells (MSCs) are currently among the best characterized adult stem cells (Majumdar et al, 1998;Pittenger et al, 1999) isolated from various tissue sources such as fat, muscle, and bone (Jiang et al, 2002b;Sottile et al, 2002;Zuk et al, 2002). These cells are able to differentiate into bone, fat, cartilage, muscle tissue, and neurons (Mackay et al, 1998;Reyes et al, 2001;Sekiya et al, 2004;Woodbury et al, 2000) and were ultimately reported in almost all cell lineages (Jiang et al, 2002;Krause et al, 2001). For these reasons, the isolation and manipulation of adult stem cells represents a promising tool for understanding tissue development and regeneration, as well as for studying the engineered repair of tissues and organs (Orlic et al, 2001;Pittenger and Martin, 2004;Wakitani et al, 2002).…”

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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Quantitative Study of Tissue-Engineered Cartilage With Human Bone Marrow Mesenchymal Stem Cells

Pang

Cui

Chen

et al. 2005

Archives of Facial Plastic Surgery

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To assess the possibility of cartilage tissue engineering using human mesenchymal stem cells (hMSCs) and to investigate the quantitative relationship between hMSCs and engineered cartilage. Design: Human mesenchymal stem cells were cultured, cryopreserved, and expanded in vitro. Surface antigens were detected by flow cytometry. In vitro chondrogenesis of hMSCs and cryopreserved hMSCs was performed. The chondrogenesis-induced hMSCs were seeded onto polyglycolic acid scaffolds, cultured in vitro for 3 weeks in chondrogenic medium, and then implanted into nude mice. The implants were harvested after 10 weeks and examined with histologic and immunochemical staining. Results: The construction of cartilages was identified grossly and histologically: 1.9 to 2.5ϫ10 7 nucleated cells were obtained from 1 mL of bone marrow, and about 1 to 2ϫ10 6 hMSCs were obtained from the primary culture. The number of hMSCs tripled at every passage and reached 1.4 to 2.8 ϫ 10 12 at passage 15. The purity of hMSCs was 95% and 98% at the primary and the fourth passages, respectively. Twenty-one days was the optimal (induction rate, 95%) induction time, with no apparent differences in induction rates among different passages. Based on our findings, hMSCs from 0.07 to 0.14 mL of bone marrow, expanded during 4 passages and induced for 21 days, would be sufficient to engineer 1 cm 2 of cartilage, 3-mm thick. Conclusion: Quantitative standards of hMSCs as seed cells for cartilage tissue engineering were established and may have value for later clinical work.

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Chondrogenic Differentiation of Cultured Human Mesenchymal Stem Cells from Marrow

Cited by 1,190 publications

References 49 publications

Adenoviral-mediated transfer of TGF-β1 but not IGF-1 induces chondrogenic differentiation of human mesenchymal stem cells in pellet cultures

Adenoviral-mediated transfer of TGF-β1 but not IGF-1 induces chondrogenic differentiation of human mesenchymal stem cells in pellet cultures

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

Quantitative Study of Tissue-Engineered Cartilage With Human Bone Marrow Mesenchymal Stem Cells

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