大鼠雪旺细胞对视网膜神经细胞成活及突起生长的影响

The Effects of Schwann Cells on Promoting Neurite Growth of Rat Retinal Neurons

:【目的】观察外周神经雪旺细胞对视网膜神经细胞成活及突起生长的影响。【方法】取出生 3～ 5d的Sprague Dawley乳鼠的视网膜组织及坐骨神经 ,用酶消化法分别得到视网膜神经细胞和雪旺细胞 ,分别和联合接种于多聚赖氨酸包被的培养皿、用含体积分数为 2 0 %胎牛血清 (fetalcalfserum ,FCS)的BME(basalmediumeagle)培养液进行培养 ,倒置相差显微镜观察细胞的生长情况 ;视网膜神经细胞和胶质细胞分别用神经丝蛋白 (neurofilament,NF)和胶质纤维酸性蛋白 (glialfibril laryacidicprotein ,GFAP)进行鉴定 ,神经节细胞用Thy1.1鉴定 ,雪旺细胞培养时用阿糖胞苷进行纯化并用S 10 0鉴定 ,扫描电镜观察共生培养的细胞。【结果】培养的视网膜细胞大部分为神经细胞 ,神经节细胞的数量较少 ;用阿糖胞苷法处理得到的绝大多数为雪旺细胞。共生培养的雪旺细胞和视网膜神经细胞可以 3种方式融合生长 :①突起与突起融合 ;②胞体与胞体融合 ;③突起与胞体融合。共生培养的视网膜神经细胞较单纯视网膜神经细胞培养的突起生长快 ,且成活的时间要长。【结论】体外培养雪旺细胞和视网膜神经细胞可以融合生长 ,前者对后者的突起生长及细胞的成活具有明显的支持和促进作用 ,这种影响作用可能与雪旺细胞分泌的神经营养因子直接导?

Objective To study the effects of Schwann cells on promoting neurite growth of rat retinal neurons in vitro. Methods The retinal tissue and sciatic nerve of newborn SD rats were prepared with enzymatic digestion to acquire retinal neurons and Schwann cells. These cells were seeded onto the poly-L-lysin covered dishes in single or in couple, cultured with BME which contain 20% fetal calf serum (FCS) and observed under phase contrast microscope. The neurons, glia cell, ganglion cells and Schwann cells were identified with NF, GFAP, Thy1 1 and S-100 respecticvely. The growth character of cells were examined with scanning electronic microscope (SEM). Results Most populations of cultured cells were retinal neurons, a few number were ganglion cells. Pure Schwann cells were obtained after Ara-c treatment. Cocultured Schwann cells and retinal neurons could grow to fuse together in following manner: ① neurite fuse neurite; ② body fuse body and ③ neurite fuse body. The neurite of retina neurons in coculture group grow faster and survive longer than that group without Schwann cells group. Conclusion The Schwann cells and retinal neurons can grow to fuse together, the former is effective to the neurite growth and survival of the latter, which suggest that the neurotrophic factors secreted by Schwann cells can enter the body of retinal neurons to make effects directly to retinal neurons survival.