[1]戴敏,张青,郑志坤,等.视网膜神经节细胞无血清上清培养液诱导大鼠视网膜干细胞的分化[J].眼科新进展,2018,38(1):035-38.[doi:10.13389/j.cnki.rao.2018.0007]
　DAI Min,ZHANG Qing,ZHENG Zhi-Kun,et al.Retinal ganglion cell-conditioned medium alters the differentiation of rat retinal progenitor cells[J].Recent Advances in Ophthalmology,2018,38(1):035-38.[doi:10.13389/j.cnki.rao.2018.0007]
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视网膜神经节细胞无血清上清培养液诱导大鼠视网膜干细胞的分化/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
38卷
期数:
2018年1期
页码:
035-38
栏目:
实验研究
出版日期:
2018-01-05

文章信息/Info

Title:
Retinal ganglion cell-conditioned medium alters the differentiation of rat retinal progenitor cells
作者:
戴敏; 张青; 郑志坤; 沈蔚
650021　云南省昆明市，云南省第二人民医院眼科 云南省眼科研究所 云南省眼科疾病防治研究重点实验室
Author(s):
DAI Min; ZHANG Qing; ZHENG Zhi-Kun; SHEN Wei
Department of Ophthalmology，the Second People’ s Hospital of Yunnan Province，Yunnan Eye Institute，Key Laboratory of Yunnan Province for the Prevention and Treatment，Kunming 650021，Yunnan Province，China
关键词:
视网膜干细胞; 视网膜神经节细胞; 视网膜神经节细胞无血清上清培养液; Nestin; Thy-1
Keywords:
retinal stem cells; retinal ganglion cells; retinal ganglion cell-conditioned medium; Nestin; Thy-1
分类号:
R774
DOI:
10.13389/j.cnki.rao.2018.0007
文献标志码:
A
摘要:
目的　探讨视网膜神经节细胞无血清上清培养液对视网膜干细胞分化的影响。方法　分离大鼠视网膜干细胞和视网膜神经节细胞；采用免疫荧光法鉴定体外培养的大鼠视网膜干细胞与视网膜神经节细胞，视网膜干细胞以Nestin抗体进行鉴定，视网膜神经节细胞以Thy-1抗体进行鉴定；以视网膜神经节细胞无血清上清培养液培养视网膜干细胞，以不加入条件培养液培养的视网膜干细胞为对照组，收集分化细胞，采用qPCR法检测Nestin、Pax6、Thy-1及Brn-3的基因表达。结果　培养的视网膜干细胞Nestin抗体染色阳性，视网膜神经节细胞Thy-1抗体染色阳性。培养视网膜干细胞72 h后，与对照组相比，无血清上清培养液组细胞Nestin和PAX6基因相对表达量降低，差异均有统计学意义（均为P<0.000 1）；Thy-1和Brn-3基因相对表达量升高，差异均有统计学意义（均为P<0.05）。结论　视网膜神经节细胞无血清上清培养液能够诱导视网膜干细胞分化为视网膜神经节样细胞。
Abstract:
Objective　To investigate the effects of retinal ganglion cell-conditioned medium on the differentiation of retinal stem cells.Methods　Retinal stem cells and retinal ganglion cells were isolated from rats，and immunofluorescence staining was applied to identify rat retinal stem cells and retinal ganglion cells with Nestin and Thy-1 antibody，respectively.Retinal stem cells were cultured in presence or absence of retinal ganglion cell-conditioned medium for 72 h，followed by detection of Nestin，PAX6，Thy-1 and Brn-3 gene expression in retinal stem cells by qPCR.Results　Isolated retinal stem cells were Nestin positive，and retinal ganglion cells were Thy-1 positive，indicating the success of isolation.Compared to retinal stem cells cultured without ganglion cell-conditioned medium，ones cultured with ganglion-conditioned medium had significantly downregulated expression of Nestin and PAX6 (both P<0.000 1)，and markedly upregulated expression of Thy-1 and Brn-3(both P<0.05).Conclusion　Retinal ganglion cell-conditioned medium can induce the differentiation of retinal progenitor cells into retinal ganglion-like cells.

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备注/Memo

备注/Memo:
昆医联合专项基金（编号：2014FB075）；云南省教育厅基金（编号：2014C046Y）
更新日期/Last Update: 2018-01-22