High throughput gene expression analysis identifies reliable expression markers of human corneal endothelial cells.

High throughput gene expression analysis identifies reliable expression markers of human corneal endothelial cells.
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高吞吐量基因表达分析确定了人角膜内皮细胞的可靠表达标记。

DOI:
10.1371/journal.pone.0067546
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发表时间:
2013
期刊:
影响因子:
3.7
通讯作者:
Colman A
Colman A
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Chng Z;Peh GS;Herath WB;Cheng TY;Ang HP;Toh KP;Robson P;Mehta JS;Colman A

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人们对通过细胞组织工程开发合适的角膜内皮移植替代品产生了极大的兴趣,这可能会缓解角膜移植材料的短缺。微创无缝合锁孔手术方案的出现,如后弹力层剥离内皮角膜移植术 (DSEK) 和后弹力层膜内皮角膜移植术 (DMEK)(仅移植内皮层而不是全层角膜),为临床应用替代内皮移植物的开发提供了进一步的动力。这项工作的一个主要挑战是缺乏这种细胞类型的特异性标记。为了鉴定在体内和体外可靠地标记角膜内皮细胞 (CEC) 的基因,我们对新鲜分离的人类 CEC(来自年轻和年长的供体)、CEC 培养物和角膜基质进行了 RNA 测序。这些角膜细胞类型的基因表达也与其他人类组织类型的基因表达进行了比较。基于高通量比较基因表达分析,我们鉴定了一组标记物: i) 在年轻供体和老年供体的 CEC 中高表达; ii) 在体内和体外在 CEC 中表达; iii)在角膜基质角膜细胞和活化的角膜基质成纤维细胞中不表达。它们是 SLC4A11、COL8A2 和 CYYR1。组合使用这组基因可以可靠地确定 CEC 细胞类型的身份。
Considerable interest has been generated for the development of suitable corneal endothelial graft alternatives through cell-tissue engineering, which can potentially alleviate the shortage of corneal transplant material. The advent of less invasive suture-less key-hole surgery options such as Descemet’s Stripping Endothelial Keratoplasty (DSEK) and Descemet’s Membrane Endothelial Keratoplasty (DMEK), which involve transplantation of solely the endothelial layer instead of full thickness cornea, provide further impetus for the development of alternative endothelial grafts for clinical applications. A major challenge for this endeavor is the lack of specific markers for this cell type. To identify genes that reliably mark corneal endothelial cells (CECs) in vivo and in vitro, we performed RNA-sequencing on freshly isolated human CECs (from both young and old donors), CEC cultures, and corneal stroma. Gene expression of these corneal cell types was also compared to that of other human tissue types. Based on high throughput comparative gene expression analysis, we identified a panel of markers that are: i) highly expressed in CECs from both young donors and old donors; ii) expressed in CECs in vivo and in vitro; and iii) not expressed in corneal stroma keratocytes and the activated corneal stroma fibroblasts. These were SLC4A11, COL8A2 and CYYR1. The use of this panel of genes in combination reliably ascertains the identity of the CEC cell type.
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发表时间: 2005-12-01
影响因子: 4.4
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DOI: 10.1186/jbiol29
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期刊: Journal of biology
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