体外培养的晶状体上皮细胞株的确定及生长、分化规律

目的建立牛、兔、人晶状体上皮细胞的体外培养，进一步认识晶状体上皮细胞生长、分化规律。方法应用组织块培养方法进行３种晶状体上皮细胞的体外培养，经Ｇｉｍｓａ染色，在倒置显微镜下对培养的晶状体上皮细胞的生长、分化规律进行观察。结果培养至６ｗｋ的人胎儿晶状体上皮细胞中有特征性的“晶状体小体”形成；牛、兔晶状体上皮细胞去分化是发生在第３代，而人晶状体上皮细胞在第４代开始出现去分化；它们传代至第８代时生长都趋于停止，出现老化表现；来自人的晶状体上皮细胞生长增殖率与年龄呈负相关关系（ｒ＝－０．９９６）。结论“晶状体小体”的形成可作为确定晶状体上皮细胞株的一项特征性依据，而体外培养的人、牛、兔晶状体上皮细胞具有相同的有限生长潜能，在相同的条件下，牛、兔晶状体上皮细胞的生长增殖速度比人晶状体上皮细胞快，但易于发生去分化；此外，人晶状体上皮细胞的生长增殖率与年龄密切相关，年龄越小，晶状体上皮细胞的生长增殖速度越快。

Study on identification and differentiation of lens epithelial cells lines in vitro

Objective To establish bovine,rabbit and human lens epithelial cells(LEC)culture and further understand the differentiation of LEC.Methods LEC from bovine,rabbit and human were cultured using tissue explant technique in vitro; The differentiation of LEC cultured from the three species was observed by Gimsa staining and a inverted microscope.Results LEC from fetus maintained in culture longer than 6 weeks spontaneously formed lens like spherules.Under identical conditions,a average doubling time of LEC from bovine,rabbit and fetus during the logarithmic phase was 24h,24.5h and 26h respectively;Dedifferentiation of cultured LEC from bovine and rabbit began after the third passage;and dedifferentiation of LEC from human occurred after the fourth passage;Cultued LEC from the three species exhibited apperant senescence at the eighth passage.The proliferative capacity of LEC from human had directly relevance to the age of donor( r =-0.996).Conclusions The formation of lens like spherules is characteristics of LEC lines in the cultured cells.Under identical conditions,the proliferative rate of LEC from bovine and rabbit is fast than that of LEC from human,but dedifferentiation of LEC from bovine and rabbit is easier than that of LEC from human;LEC from the three species exhibit similar limited growth potential.The proliferative rate of LEC from human has a inversely proportion with age.