KMID : 1004820120130040310
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Journal of Biomedical Research 2012 Volume.13 No. 4 p.310 ~ p.319
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Chondrogenesis of human umbilical cord-derived mesenchymal stem cells in vitro by TGF¥â3 and BMP6
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Park Eu-Gene
Cho Tae-Jun Kwon Soon-Keun Lee Dong-Sup Cho Jae-jin
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Abstract
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Human umbilical cord is easy to obtain because it is discarded after birth, so that ethical issues can be avoided. Chondrogenesis studies using MSCs from bone marrow, cord blood, and adipose have indicated that TGF¥â3 and BMP6 stimulate chondrogenesis. Therefore, we investigated chondrogenesis of hUC-MSCs on TGF¥â3, BMP6, and combination of the two growth factors. We initiated chondrogenesis of cells by application of physical forces to form 3D cell clusters. After initiation, we designated four experimental groups for differentiation of cells, as follows: control, 10 ng/mL TGF¥â3, 100 ng/mL BMP6, and the combination of 5 ng/mL TGF¥â3 and 50 ng/mL BMP6. For analysis of chondrogenesis, GAG contents, mRNA expression, histological analysis and immunohistochemistry (IHC) were performed. For analysis of GAG contents, GAG assay was performed and RT-PCR was performed for determination of chondrogenic markers. Histological analysis was performed through safranin O, alcian blue, and IHC was performed using collagen type I and II. GAG contents were increased 184% by TGF¥â3, 147% by BMP6, and 189% by the combination of TGF¥â3 and BMP6, compared to control. The growth factors improved collagen II and aggrecan expression; in particular, TGF¥â3 and BMP6 showed a synergistic effect, compared to only TGF¥â3 or BMP6 treated. The results of histological and IHC analysis indicated that chondrogenic differentiation in TGF¥â3 and the combination of TGF¥â3 and BMP6 showed more cartilage deposition. In conclusion, TGF¥â3 and BMP6 differentiated hUC-MSCs into chondrogenic clusters of the combination treatment of the two growth factors showed more efficient chondrogenic ability.
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KEYWORD
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human umbilical cord mesenchymal stem cell, chondrogenesis, TGF¥â3, BMP6
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