角膜缘上皮细胞体外培养、冻存和复苏的实验研究

Studies on cultures, cryopreservation and resuscitation of limbal corneal epithelial cells of rabbit in vitro

目的 建立兔角膜缘上皮细胞体外培养、冻存和复苏的方法。方法 兔角膜缘上皮细胞组织块接种培养。取第三代细胞进行冻存。于冻存后第2周,3、6个月复苏细胞。用MTT法测细胞生长曲线。结果 兔角膜缘上皮细胞体外生长良好。培养细胞AE1单克隆抗体染色阳性,PAS染色阴性。冻存细胞复苏成功。冻存细胞复苏后生长曲线良好。结论 兔角膜缘上皮细胞可以在体外培养、冻存和复苏。

Objective To establish a method of cultivating limbal corneal epithelial cells of rabbit in vitro and to study its cryopreservation and resuscitation. Methods Limbal corneal epithelial cells of rabbit were cultivated in vitro and passaged 2 times,then were identified and saved by cryopreservation. The frozen cells were resuscitated after 2 weeks, 3 monthes and 6 monthes. The cell growth curve of frozen cells were measured by MTT colorimetry. Results Limbal corneal epithelial cells of rabbits exhibited good growth during planting culture. The cultured cells stained positively for monoclonal antibody cytokeratin AE1 and negatively for PAS. The frozen cells were resuscitated successsfully. The cell growth curve of frozen cells measured by MTT colorimetry was similar to that of non-frozen cells. Conclusion The limbal corneal epithelial cells of rabbit could be cultivated, cryopreserved and resuscitated in vitro .