摘要
目的方法结果结论深低温保存培养的角膜缘上皮细胞,复苏后进行异体移植实验,为临床应用提供实验依据。将浅层角膜基质载体上培养的角膜缘上皮细胞深低温保存,同时制作兔角膜缘干细胞损伤模型,分别用新鲜培养的角膜缘上皮细胞和冷冻复苏后培养的细胞及角膜基质进行异体移植实验。角膜缘上皮细胞能成功地培养于载体上,将培养细胞保存于-196℃的液氮罐中二月,可见深低温保存前后细胞的形态结构及生物学特性无显著差异。异体移植实验显示:新鲜培养的细胞及深低温保存后的细胞异体移植后,模型兔角膜表面逐渐正常上皮化,而角膜基质组异体移植后,角膜表面仍结膜化并可见大量新生血管生长,各项检测结果与培养细胞组相比差异显著(<0.05)。以浅层角膜基质为载体培养的角膜缘上皮细胞深低温保存后仍具有上皮细胞特性并具有高增殖能力,可用以进行异体移植治疗角膜缘干细胞缺陷的角膜病。
Objective To preserve cultivated limbal epithelial ceils by cryopreservation and carry out allogeneic transplantation when revived .In order to obtain experiment evidence for clinical utilization. Methods To preserve rabbit＇s limbal corneal epithelial cells cultured on rabbit superficial layer corneal stroma by cryopreservation. 30 rabbits who undergone corneal epithelium and limbal ablation to induce limbal stem cell deficiency were randomly divided into three groups, corneal stroma group,non-frozen cells group and frozen ceils group（n=10 for each group）. To observe the each group by transplantation experiment. Results The structure and biologic viability of frozen limbal corneal epithelial cells that is stored in preservative fluid and frozen in nitrogen canister for two months did not show marked change. The allogeneic transplantation of cultured ceils on corneal stroma indicated： after the allogeneic transplantation of frozen and non-frozen subculturing cells on corneal stroma, rabbit cornea were all successfully epithelialied and corneal new vessels gradually disappeared. After transplanting corneal stroma , corneal epithelia remained defected and corneal new vessels remained invariable. Conclusion The cultivated limbal epithelial ceils on superficial layer corneal stroma still remained character of epithelial ceils and reproductive activity through refrigeration for two months .These cells growing on the carrier may be used to cure keratopathy of limbal stem cell deficiency by allogeneic transplantation of cultured cells on corneal stroma.
出处
《实用防盲技术》
2009年第4期3-7,共5页
Journal of Practical Preventing Blind
关键词
细胞培养
深低温保存
异体移植
Cell culture
Cryopreservation
Allogeneie transplantation
