Derivation of corneal endothelial cell-like cells from rat neural crest cells in vitro.
Derivation of corneal endothelial cell-like cells from rat neural crest cells in vitro.
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DOI：
10.1371/journal.pone.0042378
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Wu X
中科院分区：
文献类型：
--
作者：
Ju C;Zhang K;Wu X
本研究旨在探讨体外诱导大鼠神经脊细胞(NCC)向功能性角膜内皮细胞(CEC)样细胞分化的可行性。用成年CEC来源的条件培养液诱导大鼠NCC。免疫荧光、流式细胞仪和实时荧光定量RT-PCR检测角膜内皮分化标志物N-钙粘附素和转录因子FoxC1、Pitx2的表达。将CFDA SE标记的CEC样细胞移植到大鼠角膜内皮细胞缺乏症模型的角膜内皮中，并保持24小时的俯视姿势以允许细胞附着。术后观察长达2个月，并进行临床和组织学检查。诱导后梭形NCC转变为多角形CEC样，表达N-钙粘蛋白、FoxC1、Pitx2、闭锁带-1和钠钾泵Na+/K+ATPase。术后7、14、21、28d，实验组角膜较对照组透明，平均角膜厚度明显小于对照组。共聚焦显微镜通过聚焦和组织学分析证实，实验组中绿色荧光阳性的CEC样细胞形成了覆盖在Descemet膜上的单层。综上所述，NCC来源的CEC样细胞具有天然CEC的特征，该诱导方案为进一步从NCC诱导人CEC提供了指导。
The aim of this study was to investigate the feasibility of inducing rat neural crest cells (NCC) to differentiate to functional corneal endothelial cell (CEC)-like cells in vitro. Rat NCC were induced with adult CEC-derived conditioned medium. Immunofluorescence, flow cytometry and real time RT-PCR assay were used to detect expression of the corneal endothelium differentiation marker N-cadherin and transcription factors FoxC1 and Pitx2. CFDA SE-labeled CEC-like cells were transplanted to the corneal endothelium of a rat corneal endothelium deficiency model, and an eye-down position was maintained for 24 hours to allow cell attachment. The animals were observed for as long as 2 months after surgery and underwent clinical and histological examination. Spindle-like NCC turned to polygonal CEC-like after induction and expressed N-cadherin, FoxC1, Pitx2, zonula occludens-1 and sodium-potassium pump Na+/K+ ATPase. The corneas of the experimental group were much clearer than those of the control group and the mean corneal thickness in the experimental group was significantly less than in the control group7, 14, 21 and 28 days after surgery. Confocal microscopy through focusing and histological analysis confirmed that green fluorescence-positive CEC-like cells formed a monolayer covering the Descemet’s membrane in the experimental group. In conclusion, CEC-like cells derived from NCCs displayed characters of native CEC, and the induction protocol provides guidance for future human CEC induction from NCC.
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影响因子：
4
作者：
Jiang, Xiaohua;Gwye, Ynnez;Lawlor, Elizabeth R.
通讯作者：
Lawlor, Elizabeth R.
影响因子：
21.3
作者：
Fernandes, KJL;McKenzie, IA;Miller, FD
通讯作者：
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影响因子：
4.4
作者：
Koh, Shay-Whey M.;Chandrasekara, Krish;Rutzen, Allan R.
通讯作者：
Rutzen, Allan R.
DOI：
10.1073/pnas.76.1.464
发表时间：
1979-01-01
影响因子：
11.1
作者：
GOSPODAROWICZ, D;GREENBURG, G;ALVARADO, J
通讯作者：
ALVARADO, J
影响因子：
14
作者：
Pang, Kunpeng;Du, Liqun;Wu, Xinyi
通讯作者：
Wu, Xinyi