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Efficient and Scalable Directed Differentiation of Clinically Compatible Corneal Limbal Epithelial Stem Cells from Human Pluripotent Stem Cells
Published on: October 24, 2018
[A pilot study on transdifferentiation of skin stem cell in reconstructing corneal epithelium]
Bing Huang1, Zhi-chong Wang, Jian Ge
1Zhongshan Ophthalmic Center of Sun Yet-sen University, GuangZhou 510060, China.
Objective:
To observe the differentiation and development of skin stem cells on corneal stroma and to discuss the possibility of reconstructing corneal epithelium with skin stem cells.
Methods:
Pieces of human and rabbit skin were obtained during operation. Rabbit eye balls were taken, and pieces of corneal stroma without epithelium were prepared. Skin stem cells from the rabbit skin and human skin were cultured. The human skin stem cells of the first generation to 4th generation were implanted on the rabbit corneal stroma and cultured. Three rabbits underwent autotransplantation of the rabbit skin stem cells of the first generation to 4th generation on the pieces of corneal stroma with the superficial lamina removed and then fed for 100 approximately 114 days. Another 3 rabbits underwent allotransplantation of the rabbit skin stem cells of first to 4th generation on the pieces of corneal stroma with the superficial lamina removed and then fed for 100 days. Then the rabbits were killed and their eye balls taken out. The rabbit corneas implanted with human or rabbit epithelial cells and the rabbit corneas with the autogeneous or heterogeneous epithelial cells were sliced and underwent immunohistochemistry with human AE5 antibody corresponding to the specific surface marker keratin K3/K12 common to humankind and rabbit, and human epithelial cell keratin K-19 monoclonal antibody.
Results:
Since the 3(rd) day of transplantation the transplanted human epithelial cells formed multiplayer and were human AE5 antibody and human K19 monoclonal antibody positive. The autotransplanted corneas remained basically transparent without obvious vascular hyperplasia till the cornea specimens were taken. Histological examination showed intact multiplayer epithelium and immunohistochemistry showed human AE5 positive. The allotransplanted\rabbit corneas showed congestion since the 9(th) day. Histological examination showed that the corneas were nor so transparent as the autotransplanted ones and the epithelium was nor intact with a lot of lymphocyte infiltration.
Conclusion:
Corneal epithelium can be reconstructed from skin stem cell, which may be an alternative for constructing autogeneous bioengineered corneas.
