P63标记角膜缘干细胞的可行性研究

Evaluation of P63 in Identification of Corneal Epithelial Stem Cell

目的探讨p63蛋白(P63)作为角膜缘干细胞阳性标记物的可行性和意义,为研究和鉴定角膜缘干细胞存活、增殖和分化情况提供依据。方法取正常新西兰兔角膜全层标本5份(包括角膜缘和结膜),同时取角膜缘干细胞移植后3个月上皮生长良好的角膜全层标本和角膜失代偿标本各5份。免疫组织化学SP三步法检测P63在不同组标本供体干细胞的表达情况。结果P63在角膜缘干细胞核内呈高表达,而短暂扩充细胞虽然也具备一定的增殖和分化能力,但是P63表达较少。角膜中央和结膜呈阴性表达。结论P63可以特异性地标记角膜缘干细胞,对角膜缘干细胞的研究和鉴定及其在眼表重建的应用具有一定的价值。

Objective To investigate the feasibility and significance of p63 antibody as a positive marker of corneal epithelial stem cells. Methods Fifteen New-Zealand rabbits were divided into three groups： the autograft group, the allograft group and the control group. Corneal epithelial stem cells were obtained by autograft and allograft limbal stem cell transplantation. Rabbits were sacrificed after three months. The corneal samples were collected and fixed with 10% formalin and performed sections for examination by immunohistochemistry method（SP approach）. Corneal tissue were analyzed by immunohistochemistry method（SP approach）. Results p63 was is abundantly expressed in nucleus of limbal stem cells. The immediately next generation expressed p63 also, but was less than limbal stem cell. The central part of cornea and conjunctiva were negative. Conclusion The identification of p63 as a keratinocyte stem cell marker will be of practical importance for the identification of limbal stem cells in clinical application of cornea reconstruction.