人视网膜中神经干细胞样细胞的分离和体外培养

目的 分离培养发育基本成熟的人视网膜中神经干细胞样细胞。方法 取0～3岁猝死婴幼儿视网膜细胞，在干细胞选择性培养基中培养，观察原代和传代细胞的形态和增殖情况，并对其诱导分化。通过免疫细胞化学方法检测Nestin、MAP-2和GFAP的表达。结果 原代细胞可形成悬浮生长的致密球状细胞团，传代后能形成新的细胞球。部分细胞球表达Nestin，传代后阳性率增加。分化后细胞形态多样，每一团细胞相对一致，部分细胞表达MAP-2或GFAP。结论 0～3岁人视网膜中存在一种表现出神经干细胞特性的细胞。

Isolation and culture of cells with neural stem cell characters from infant human retina

Objective To isolate the neural stem cell-like cells from infant human retinas. Methods Ten healthy retinas were gotten from died infants,and the retina cells were cultured in special medium to obtain neural stem cells.The growth and proliferation of primary and passage cells were observed,and differentiation of cultured cells was induced.An immunocytochemial method was used to identify the expression of Nestin,MAP-2 and GFAP. Results The primary cultured retinal cells could form a condensing sphere-shape cell cluster,and then they separated from the wall of culture vessel and suspended in culture medium.Daughter spheres could form after the primary sphere were digested into single or small cluster.The expression of Nestin could be detected both in the primary cell spheres and daughter ones in part,and the numbers of positive cells were increased in daughter spheres.The differentiated cells displayed various shapes and expressed MAP-2 or GFAP. Conclusion The infant human retina has some cells with neural stem cell characters.