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Study on method of directional differentiation of human induced pluripotent stem cells towards mesenchymal stem cells

HU Guo-wen1, LI Qing2, NIU Xin2, HU Bin2, LIU Juan1, LANG Hai-li4, SHEN Xiao-li1, WANG Yang2, DENG Zhi-feng1,3   

  1. 1.Department of Neurosurgery, the Second Affiliated Hospital of Nanchang University, Nanchang 330006, China; 2.Institute for Microsurgery of Extremities,the Sixth People's Hospital, Shanghai Jiao Tong University, Shanghai 200233, China; 3.Department of Neurosurgery, the Sixth People's Hospital, Shanghai Jiao Tong University, Shanghai 200233, China; 4.Department of Anesthesiology, the Second Affiliated Hospital of Nanchang University, Nanchang 330006, China
  • Online:2014-10-28 Published:2014-10-28
  • Supported by:

    National Natural Science Foundation of China, 81272170; 81472152

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Abstract:

Objective To develop an efficient and simple method to induce the differentiation of human induced pluripotent stem cells (hiPS) towards mesenchymal stem cells (MSCs). Methods MSCs induction medium was used to induce iPS towards MSCs. The cell morphology was observed under the inverted microscope during the course of induction. The expressions of surface markers of iPS and iPS-MSCs were detected by the flow cytometry. The expressions of stemness genes such as NANOG, OCT-4, and MSX-1 in cells during the course of differentiation were detected by the RT-PCR. The osteogenesis, adipogenesis, and chondrogenesis were used to identify their tri-lineages differentiation pluripotency. Results The iPS gradually grew outward after being induced and MSCs gradually became fusiform in Passage 4. The iPS-MSCs were positive to CD29, CD44, CD73, CD90, and CD105 and negative to CD34, CD45, and HLA-DR. The expressions of NANOG and OCT-4 gradually decreased during the course of differentiation and NANOG and OCT-4 were not expressed in iPS-MSCs. The expression of MSX-1 increased to a high level after being induced for 4 d and maintained afterwards. The iPS-MSCs had abilities of osteogenesis, adipogenesis, and chondrogenesis. Conclusion An efficient and simple method is developed for directly inducing iPS towards functional iPS-MSCs, which provides the technical basis for further research and application of iPS-MSCs.

Key words: induced pluripotent stem cells, mesenchymal stem cells, cell differentiation