人泪腺上皮细胞的体外培养及鉴定

In vitro culture and identification of humanh lacrimal gland epithelial cells

目的 探讨人泪腺上皮细胞原代体外培养、鉴定、冻存和复苏的方法。方法 应用组织块培养法和混合消化液培养法对健康猝死成年人泪腺上皮细胞进行体外原代培养 ,应用免疫组织化学染色方法对培养有第 2代上皮细胞进行Pan keratin蛋白染色 ,以进行鉴定 ,取第 2、4代细胞进行冻存 ,1个月后取出复苏。结果 组织块培养法和混合消化液培养法均可获得较纯的泪腺上皮细胞 ,以组织块培养法接种的细胞早期生长较快 ,但第一代以后两种方法所获得的细胞生长无明显差别。两种方法所获取的泪腺上皮均为贴壁生长 ,未能形成生理状态下泪腺的囊腔结构 ,亦未见分泌小泡的形成。培养的泪腺上皮免疫组化染色Pan keratin蛋白染色阳性。经冻存和复苏 ,细胞保持良好活性。结论 组织块培养法和混合消化液培养法均可获得较纯的泪腺上皮细胞 。

Objective To establish the in vivo culture method of human lacrimal gland(LG) epithelial cells and the identification, storage and revivification of the cells Methods The LG epithelial cells were obtained by tissue culture or enzymatic digestion The expression of Pan keratin was detected in the second generation of epithelial cells using immunohistochemistry Store the 2nd and 4th generatin of LG cells by freezing and then revivificate them one month later Results Pure LG epithelial cells can be obtained by these two methods The cells growed faster in the early stage However, the cells were of no differen after the 1st generation The cells growed well on plastic culture flask, but could not form the lumens or acinar like structure The cultured LG epithelial cells showed positive staining of Pan karatin After freezed storage and revivification, the LG epithelial cells growed well Conclusion LG epithelial cells obtained by tissue cuture and enzymatic digestion are relatively pure, but cannot form normal lumen structure