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Treatment with retinoic acid and lens epithelial cell-conditioned medium in vitro directed the differentiation of pluripotent stem cells towards corneal endothelial cell-like cells

  • Authors:
    • Ping Chen
    • Jun‑Zhao Chen
    • Chun‑Yi Shao
    • Chuan‑Yin Li
    • Yi‑Dan Zhang
    • Wen‑Juan Lu
    • Yao Fu
    • Ping Gu
    • Xianqun Fan

  • View Affiliations

  • Published online on: December 3, 2014     https://doi.org/10.3892/etm.2014.2103
  • Pages: 351-360

  • Copyright: © Chen et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) have extensive self‑renewal capacity and the potential to differentiate into all tissue‑specific cell lineages, including corneal endothelial cells (CECs). They are a promising prospect for the future of regenerative medicine. The method of derivation of CECs from ESCs and iPSCs, however, remains to be elucidated. In this study, mouse ESCs and iPSCs were induced to differentiate into CECs using CEC embryonic development events as a guide. All‑trans retinoic acid (RA) treatment during the embryoid body (EB) differentiation step was used to promote neural crest (NC) cell differentiation as first step and was followed by a second induction in CEC‑ or lens epithelial cell (LEC)‑conditioned medium (CM) to ultimately generate CEC‑like cells. During the corresponding differentiation stages, NC developmental markers and CEC differentiation markers were detected at the protein level using immunocytochemistry (ICC) and at the mRNA level by reverse transcription‑quantitative polymerase chain reaction (RT-qPCR). During the first stage, the data indicated that 4 days of treatment with 1 µM RA starting on day 4 of EB formation favored NC cell differentiation and that plating on gelatin‑coated plates led to cell migration out of the EBs. The second‑stage differentiation results showed that the CM, particularly the LEC‑CM, enhanced the yield of polygonal cells with CEC‑specific marker expression shown by ICC and RT-qPCR. This study demonstrates that mouse ESCs and iPSCs were induced and expressed CEC differentiation markers when subjected to a two‑step inducement process, suggesting that they are a promising resource for corneal endothelium failure replacement therapy in the future.
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February-2015
Volume 9 Issue 2

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Spandidos Publications style
Chen P, Chen JZ, Shao CY, Li CY, Zhang YD, Lu WJ, Fu Y, Gu P and Fan X: Treatment with retinoic acid and lens epithelial cell-conditioned medium in vitro directed the differentiation of pluripotent stem cells towards corneal endothelial cell-like cells. Exp Ther Med 9: 351-360, 2015.
APA
Chen, P., Chen, J., Shao, C., Li, C., Zhang, Y., Lu, W. ... Fan, X. (2015). Treatment with retinoic acid and lens epithelial cell-conditioned medium in vitro directed the differentiation of pluripotent stem cells towards corneal endothelial cell-like cells. Experimental and Therapeutic Medicine, 9, 351-360. https://doi.org/10.3892/etm.2014.2103
MLA
Chen, P., Chen, J., Shao, C., Li, C., Zhang, Y., Lu, W., Fu, Y., Gu, P., Fan, X."Treatment with retinoic acid and lens epithelial cell-conditioned medium in vitro directed the differentiation of pluripotent stem cells towards corneal endothelial cell-like cells". Experimental and Therapeutic Medicine 9.2 (2015): 351-360.
Chicago
Chen, P., Chen, J., Shao, C., Li, C., Zhang, Y., Lu, W., Fu, Y., Gu, P., Fan, X."Treatment with retinoic acid and lens epithelial cell-conditioned medium in vitro directed the differentiation of pluripotent stem cells towards corneal endothelial cell-like cells". Experimental and Therapeutic Medicine 9, no. 2 (2015): 351-360. https://doi.org/10.3892/etm.2014.2103