人角膜缘干细胞在体外不同载体上培养的实验研究

目的 寻找人角膜缘干细胞体外培养的载体。方法 人角膜缘干细胞原代培养单细胞层传代到羊膜、卵壳膜、聚乳酸膜、藻酸盐凝胶4种载体和6种培养板中，倒置显微镜观察、记录细胞生长情况。对6孔板传代细胞行生物特性检测，对载体上培养细胞行组织学检测。结果 6孔板中传代细胞对AE－5，4G10．3表达阳性。羊膜、卵壳膜和聚乳酸膜上有细胞生长，7天左右形成单层；藻酸盐凝胶载体上未见细胞生长。结论 传代细胞具有角膜缘干细胞生物特性；羊膜、卵壳膜和聚乳酸膜适合角膜缘干细胞传代培养。

Experimental investigation of human corneal limbal stem cells vector culture invitro

Objective To select ideal vectors for human corneal stem cells(Scs) subculture in vitro. Methods The primary intact SCs sheet released with the Dispase were subcultured to six-well culture plate as well as to four different vectors: amniotic membrane,ovalshell membrane,polylactic membrane and alginate,observing cells under inverted culture microscope every day. Chararteristics of subcultured SCs were detected by immunohistochemical and histological methods. Results Cells subcultured to vectors of amniotic membrane, polylartic membrane and ovalshell membrane could adhere and grow within 24 hours,forming comment monolayer in seven days. However,cells on alginate could not survive. Subcultured SCs in six-well plate were positive to monoclonal antibody AE-5 and 4 G10. 3. Conclusions SCs subcultured in vitro still maintain the characteristics of SCs. Amniotic membrane, polylactic membrane and ovalshell membrane are suitable for corneal SCs subculture and is safe vectors for cell culture and transplantation.