[1]李岩,贺新征,李军,等.睫状缘色素细胞与骨髓间充质干细胞体外共培养构建视网膜干细胞的初步研究[J].眼科新进展,2016,36(1):019-22.[doi:10.13389/j.cnki.rao.2016.0006]
　LI Yan,HE Xin-Zheng,LI Jun,et al.Preliminary study of retinal stem cells fabrication by direct co-culture of ciliary margin pigment cells and bone marrow mesenchymal stem cells[J].Recent Advances in Ophthalmology,2016,36(1):019-22.[doi:10.13389/j.cnki.rao.2016.0006]
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睫状缘色素细胞与骨髓间充质干细胞体外共培养构建视网膜干细胞的初步研究
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
36卷
期数:
2016年1期
页码:
019-22
栏目:
实验研究
出版日期:
2016-01-05

文章信息/Info

Title:
Preliminary study of retinal stem cells fabrication by direct co-culture of ciliary margin pigment cells and bone marrow mesenchymal stem cells
作者:
李岩; 贺新征; 李军; 倪芳芳; 孙青青
315040　浙江省宁波市，解放军第113医院
Author(s):
LI Yan; HE Xin-Zheng ;  LI Jun ; NI Fang-Fang ; SUN Qing-Qing
Department of Ophthalmology,the ll3th Hospital of PLA,Ningbo 315040,Zhejiang Province.China
关键词:
睫状缘色素细胞; 骨髓间充质干细胞; 视网膜干细胞; 细胞共培养
Keywords:
ciliary margin pigment cell; bone marrow mesenchymal stem cell;  retinal ganglion cell;  co-culture
DOI:
10.13389/j.cnki.rao.2016.0006
文献标志码:
A
摘要:
目的　探索睫状缘色素细胞（ｐｉｇｍｅｎｔｅｄｃｅｌｌｓｆｒｏｍｔｈｅｃｉｌｉａｒｙｍａｒｇｉｎ，ＰＣＭ）与骨髓间充质干细胞（ｂｏｎｅｍａｒｒｏｗｍｅｓｅｎｃｈｙ-ｍａｌｓｔｅｍｃｅｌｌｓ，ＢＭ）体外共培养构建视网膜干细胞的可行性。方法　按照组织贴壁培养方法分别分离培养原代大鼠ＢＭ和ＰＣＭ，再将二者进行直接共培养（１２），观察细胞生长情况；ＭＴＴ方法进行增殖能力检测；在促神经分化诱导液中诱导２１ｄ后行免疫荧光染色，观察视网膜干细胞相关分子标记物包括视杆细胞Ｒｈｏ１Ｄ４、双极神经元ＣＨＸ１０和Ｍüｌｌｅｒ胶质细胞１０Ｅ４的表达。结果　原代分离培养的两种细胞生长状态良好；共培养后细胞总体的增殖活性虽然显著低于单纯ＢＭ，但比单纯ＰＣＭ有了一定提升；诱导分化后单纯ＰＣＭ视网膜干细胞相关分子标记物表达阳性率显著高于ＢＭ，而共培养后的细胞表达阳性率显著高于单纯ＰＣＭ和ＢＭ。结论　采用ＢＭ与ＰＣＭ共培养能够获得大量表达视网膜干细胞相关分子标记物的细胞群，有望成为视网膜干细胞来源，用于视神经损伤修复。
Abstract:
Objective To explore the availability of fabrication retinal stem cells (RSC) in vitro by co-culture of pigmented cells from the ciliary margin ( PCM) and bone marrow mesenchymal stem cells ( BM) . Methods Primary PCM and BM were isolated according to previous reports. Co-culture was performed by directly combine BM and PCM at the volume ratio of I : 2. The growth was observed by inverted microscope , while the proliferation was monitored by MTT assay. After neural induction for 21 days ,RGC related markers of rod photoreceptor ( RhoID4 ) : bipolar neurons ( CHXIO) and Muller glia ( IOE4 ) were detected by immunofluorescent staining. Results The isolated cells grew in good condition. The proliferation rate of co-cultured cells was lower than BM , but higher than PCM. After induction. RGC related markers expression in PCM were significantly higher than BM. while the co-cultured cells were much higher than PCM and BM. Conclusion Direct co-culture of BM and PCM can acquire abundant RSC , and may be a promising cell source for damaged optical neuron repair.

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更新日期/Last Update: 2016-01-04