Huang Bing, Wang Zhi-chong, Ge Jian, Chen Xi-gu, Liu Jing-bo, Fan Zhi-gang, Tao Jing, Liu Bing-qian, Guo Fen-fen
Zhongshan Ophthalmic Center of Sun Yet-sen University, GuangZhou 510060, China.
Zhonghua Yi Xue Za Zhi. 2004 May 17;84(10):838-42.
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.
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.
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.
Corneal epithelium can be reconstructed from skin stem cell, which may be an alternative for constructing autogeneous bioengineered corneas.
观察皮肤干细胞在角膜基质上的分化与发育，探讨用皮肤干细胞重建角膜上皮的可能性。
手术中获取人及兔皮肤组织。摘取兔眼球，制备无上皮的角膜基质片。培养兔皮肤和人皮肤的干细胞。将第1代至第4代人皮肤干细胞植入兔角膜基质并培养。3只兔将第1代至第4代兔皮肤干细胞自体移植到去除浅层的角膜基质片上，饲养约100至114天。另外3只兔将第1代至第4代兔皮肤干细胞异体移植到去除浅层的角膜基质片上，饲养100天。然后处死兔子，取出眼球。将植入人或兔上皮细胞的兔角膜以及自体或异体上皮细胞的兔角膜切片，用对应人类和兔共有的特异性表面标志物角蛋白K3/K12的人AE5抗体以及人上皮细胞角蛋白K - 19单克隆抗体进行免疫组织化学检测。
移植后第3天起，移植的人上皮细胞形成多层，人AE5抗体和人K19单克隆抗体呈阳性。自体移植的角膜直至取材时基本保持透明，无明显血管增生。组织学检查显示有完整的多层上皮，免疫组织化学显示人AE5阳性。异体移植的兔角膜自第9天起出现充血。组织学检查显示角膜不如自体移植的透明，上皮不完整，有大量淋巴细胞浸润。
皮肤干细胞可重建角膜上皮，这可能是构建自体生物工程角膜的一种替代方法。
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