影响大鼠胚胎视网膜祖细胞体外增殖的两个因素

Two factors that influence proliferation of retinal progenitor cells from rat embryo in vitro

目的探讨在体外培养条件下影响大鼠胚胎视网膜祖细胞增殖的因素。方法取第18天大鼠胚胎,用无血清培养技术扩增视网膜祖细胞,以形成悬浮生长的细胞球,经免疫细胞化学、扫描电镜与透射电镜技术鉴定细胞球中已分化的细胞,并观察不同细胞接种密度对祖细胞增殖率的影响。结果①细胞球中出现分化细胞形态和NF及Nestin阳性细胞;②提高接种细胞密度,祖细胞增殖率增加。结论①此期的祖细胞分裂产生分化细胞,导致祖细胞增殖能力差;②接种细胞密度影响祖细胞增殖率。

Objective To investigate two factors influencing proliferation of retinal progenitor cells (RPCs) from rat embryo in vitro. Methods RPCs from 18-day rat embryo (E18) proliferated to form neural spheres by using serum-free culture. Differentiated cells among them were identified by using immunocytochemistry, scanning and transmission electron microscope;moreover, the effect of different cell density to the clone efficiency of RPCs was observed and compared. Results ①Neural spheres have the morphology of differentiated cells, and NF positive and Nestin positive cells. ②The increase of cell density resulted in the rise of clone efficiency of RPCs. Conclusions ①Some RPCs divided in vitro and produced differentiated cells, so RPCs from E18 proliferated badly. ②The clone efficiency of RPCs is affected by cell density.