Effects of transforming growth factor β3, hyaluronic acid, and parathyroid hormone-related protein on chondrogenic differentiation of human umbilical cord mesenchymal stem cells
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(1. Laboratory Animal Center, Southern Medical University, Guangzhou 510000, China.2. Southern Theater Air Force Hospital, Guangzhou 510000. 3. Wuyi University, Jiangmen 529020)

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R-33

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    Abstract:

    Objective To investigate the effects of transforming growth factor (TGF)-β3, hyaluronic acid (HA), and parathyroid hormone-related protein ( PTHrP) on chondrogenic differentiation of human umbilical cord mesenchymal stem cells and to optimize the combination of culture conditions. Methods Human umbilical cord mesenchymal stem cells were isolated and cultured in vitro for morphological examination, surface antibody detection, and adipogenic and chondrogenic differentiation. Different combinations of cytokines were added to P3 generation cells for chondrogenic differentiation as follows: TGF-β3 on days 1-21 (G1), TGF-β3 on days 1-28 (G2), TGF-β3 on days 1-21 + HA on days 1-21 (G3), TGF-β3 on days 1-28 + HA on days 1-28 (G4), TGF-β3 on days 1-21 + HA on days 1-21 + PTHrP on days 14-21 (G5), TGF-β3 on days 1-28 + HA on days 1-28 + PTHrP (G6) on days 14-28, and TGF-β3 on days 1-28 + HA on days 1-28 + PTHrP on days 21-28 (G7). Results The expression levels of COL2α1, COL10α1, SOX9, and ACAN were higher in the TGF-β3 and TGF-β3/ HA groups at 28 than 21 days of culture.In addition, the expression levels of COL2α1, SOX9, and ACAN were higher and that of COL10α1 was lower in the TGF-β3/ HA/ PTHrP group. When cultured at the same times, the expression levels of COL2α1, SOX9, and ACAN were higher in the TGF-β3/ HA group than TGF-β3 group, and the expression levels of COL2α1, ACAN, and SOX9 were higher and that of COL10α1 was lower in the TGF-β3/ HA/ PTHrP group than TGF-β3/ HA group. The effect of PTHrP addition for 2 weeks was significantly more obvious than that for 1 week ( P <0. 05). The results of hematoxylin-eosin and alcian blue staining were consistent with the results of real-time quantitative polymerase chain reaction. Conclusions TGF-β3, HA, and PTHrP can promote chondrogenic differentiation to different extents. When TGF-β3 and HA were combined for 28 days and PTHrP was added on the 14th day (G6), this combination promotes best the chondrogenic differentiation of human umbilical cord mesenchymal stem cells and inhibits the cell hypertrophy.

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History
  • Received:March 04,2019
  • Revised:
  • Adopted:
  • Online: November 07,2019
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