以羊膜为底物培养鸡角膜缘上皮细胞的实验研究

The culture of chicken corneal limbal epithelial cells on amnion basement membrane

目的 为眼表面重建术提供良好的移植材料。方法 用胰酶及EDTA消化法去除羊膜上皮将消化、离心获得的鸡角膜缘上皮细胞调成 1× 10 7/mlDMEM悬液 ,接种在 6孔培养板底的羊膜基底膜面 ,放入 3 7℃ ,5 %CO2 孵箱培养。结果 48h羊膜上培养细胞形成单层 ,较培养板上培养的同种细胞小 ,细胞表面的微绒毛及侧面足状突丰富 ,立体感明显。结论 以羊膜基底膜为底物培养的鸡角膜缘上皮细胞可以着床生长并形成完整的单层上皮 ,为眼表重建使用羊膜及基底膜面培养的角膜上皮细胞提供了实验材料。

ObjectiveTo provide the cultured quality materials for clinical transplantation.MethodsRemoval of amniotic membrane epithelium was carried on with the digestion method of trypsine/EDTA.Suspension of chicken corneal limbal epithelial cells in DMEM(Dulbecco’s Medified Eagle’s Medium)were seeded on the amnion basement membrane(ABM)laid a flat on the basement of culture well at a density of 1×10 7 cells/ml,then,were put into the incubator at 37℃ in a humidified atmosphere containing 5% CO 2.ResultsA monolayer of cultured chicken corneal limbal epithelial cells was formed in 48 h.Stereo views of these cells on AM were obvious;they were smaller,the numbers of microvilli of their surfaces and footprocesses on the sides of them were larger than those of cultured cells on the culture plate at the same time.ConclusionThe cultured chicken corneal limbal epithelial cells on ABM can form into monolayer,which provids experimental materials for reconstrution of ocular surface.It is proved that AM can promote the growth of cultured corneal limbal epithelial cells in vitro.