体外诱导室管膜下区神经干细胞分化为视网膜神经细胞样细胞的研究

Induced differentiation of neural stem cells from subependymal zone into retinal cells in vitro

目的 研究室管膜下区细胞的体外培养及其诱导分化情况。方法 取新生 0～ 3d的SD乳鼠室管膜下区细胞进行体外培养 ,同时培养视网膜神经细胞。收集视网膜细胞培养上清液 ,配制成条件分化液 ,将培养的室管膜下区细胞接种于视网膜细胞条件分化液中 ,并对诱导的细胞进行免疫荧光鉴定。结果 体外培养的室管膜下区细胞在无血清的培养基中生长良好 ,接种于SD乳鼠视网膜细胞条件分化液中的室管膜下区细胞可分化为视网膜神经样细胞。应用Thy1 1单克隆抗体行免疫荧光检查 ,细胞呈阳性反应。结论 在一定条件下室管膜下区细胞能够于体外培养存活 ,视网膜细胞条件分化液可定向诱导室管膜下区细胞分化为视网膜神经节细胞样细胞 ,体外模拟眼内环境行室管膜下区细胞诱导 ,将为进一步的眼内移植提供理论依据。

Objective To study the cultivation and differentiation of neural stem cells (NSC) in vitro and observe the effects of retinal cell-conditioned medium on the differentiation of NSC. Method Cells from subependymal zone of postnatal 0-3 days SD rat were isolated and cultured in vitro. Retinal cells of SD rat were cultured simultaneously. The supernatants (conditioned medium, SDR-CM) of cultured retinal cells were collected and used for the cultivation of the NSC from subependymal zone. Immunofluorescence method and anti-Thy1.1 antibodies were used to identify the cells derived from the NSC. Results Cultured NSC grew well in the serum-free culture medium. Cultured subependymal cells in the SDR-CM could be differentiated to the retinal cells. Some cells were stained positively with anti-Thy1.1 antibodies. Conclusion These results showed that cultured NSC could survive well in vitro. SDR-CM can induce the NSC to differentiate to the retinal cells. The present study was designed to simulate the microenvironment of the eyes and to induce the differentiation of the NSC. Our in vitro model system may provide theoretical basis for the intraocular transplantation of retinal neurons.