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Experimental study on differentiating human induced pluripotent stem cells towards renal cells in vitro

HUANG Jia-hui1,2, NIU Xin1, HU Bin1, ZHOU Shu-min1, LI Qing1, GUO Shang-chun1, WANG Yang1   

  1. 1.Shanghai Institute for Microsurgery of Extremities, the Sixth People's Hospital, Shanghai Jiao Tong University, Shanghai 200233, China; 2.Department of Clinical Laboratory, the Sixth Affiliated Hospital of Sun Yat-Sen University, Guangzhou 510655, China
  • Online:2014-07-28 Published:2014-08-11
  • Supported by:

    National Nature Science Foundation of China, 81170640, 30960385

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Abstract:

Objective To investigate the method of differentiating human induced pluripotent stem (iPS) cells towards renal cells in vitro and to provide the experimental basis for the application of stem cells in kidney regeneration. Methods The normal human iPS cells were treated by cytokines such as Activin-A, BMP7, hVEGF, b-FGF, lithium, and retinoic acid (RA) in renal epithelial cell growth medium (REGM) for inducing the differentiation towards renal cells. After 14, 21, and 28 d of differentiation, the expressions of proteins relevant to the renal development, i.e. Bry, Pax2, AQP1, and E-cad, were detected by the immunofluorescence staining and Real-Time PCR. The expressions of pluripotent genes of Nanog and OCT4 of each differentiation stage were measured by the RT-PCR. Results After being induced by the medium for 14, 21, and 28 d, the results of immunofluorescence staining showed that the differentiated cells expressed the proteins of Bry, Pax2, AQP1, and E-cad, while undifferentiated iPS cells did not express. The results of Real-Time PCR indicated that the expressions of Bry, Pax2, AQP1, and E-cad of differentiated cells were about 4, 30, 37, and 25 times higher than those of undifferentiated iPS cells (P<0.05). The expressions of pluripotent genes of Nanog and OCT4 of differentiation stages decreased gradually (P<0.05). Conclusion Human iPS cells can be efficiently induced and differentiate towards renal cells by cytokines in renal epithelial cell growth medium.

Key words: cytokines, differentiation, induced pluripotent stem cells, renal cells