摘要
目的探讨乳鼠视网膜细胞对大鼠骨髓间充质干细胞(BMSCs)的诱导分化作用。方法从出生8d的大鼠骨髓中分离培养BMSCs,同时培养乳鼠视网膜神经细胞。在Transwell双层培养体系下,利用乳鼠视网膜细胞诱导BMSCs,诱导后细胞经RT-PCR及免疫荧光鉴定。结果加入乳鼠视网膜细胞共培养第7d有神经元样细胞形成,经nestin、NF、β-IIITubulin、Thy1.1行免疫组织化学、RT-PCR、免疫荧光鉴定,细胞呈阳性反应。结论BMSCs在体外培养条件下,经过新生乳鼠视网膜细胞诱导,可以分化为视网膜神经节样细胞。
Objective Previous studies had shown that bone marrow mesenchymal cells（BMSCs） could be induced into photoreceptors. The research of neurology verified that the development of retina depends upon the time phase after BMSCs transplantation. This study was undertaken to explore the inducing effects of retinal cells of rat neonates on differentiation of bone marrow mesenchymal cells（BMSCs） into RGC-like cells. Methods The retinal cells from 3-day Wistar rats and BMSCs from postnatal 8 days of rat neonates were isolated,digested by trypsin and cultured in the DMEM/F12 containing 10% fetal bovine serum separately. The BMSCs were induced by retinal cells of rat neonates in transwell. The expression of NF, nestin, β-Ⅲ Tubulin were detected, and immunoflurorescence method and RT-PCR were used for the identification of the cells transdifferentiation. Results On the 7th day after adding retinal cells of rat neonates, the neural cell showed modality-like in shape and presented the positive reaction for CD71 , CD44 and negative reaction for CD45, CD34. The differentiated retinal cells showed round in shape with process and branch and network-like connect and presented the positive staining for nestin, NF, β-Ⅲ Tubulin and Thyl. 1 antibody. However,the non-neutron-like cells presented the polygon and spindle-like in shape with the absent reaction for nestin,NF,β-Ⅲ Tubulin and Thyl. 1 antibody. Conclusion BMSCs can be induced into RGC-like cells by retinal cells of rat neonates.
出处
《眼科研究》
CAS
CSCD
北大核心
2008年第3期174-178,共5页
Chinese Ophthalmic Research
基金
国家自然科学基金资助(30471858)
