慢周转角膜缘上皮细胞的特征鉴定出潜在的干细胞标志物。

Characterization of slow cycling corneal limbal epithelial cells identifies putative stem cell markers.

机构信息

University of Illinois, Chicago, USA.

Weill Cornell Medical College, New York, USA.

出版信息

Sci Rep. 2017 Jun 19;7(1):3793. doi: 10.1038/s41598-017-04006-y.

In order to identify reliable markers of corneal epithelial stem cells, we employed an inducible transgenic "pulse-chase" murine model (K5Tta × TRE-H2BGFP) to localize, purify, and characterize slow cycling cells in the cornea. The retention of GFP labeling in slowly dividing cells allowed for localization of these cells to the corneal limbus and their subsequent purification by FACS. Transcriptome analysis from slow cycling cells identified differentially expressed genes when comparing to GFP faster-dividing cells. RNA-Seq data from corneal epithelium were compared to epidermal hair follicle stem cell RNA-Seq to identify genes representing common putative stem cell markers or determinants, which included Sox9, Fzd7, Actn1, Anxa3 and Krt17. Overlapping retention of GFP and immunohistochemical expression of Krt15, ΔNp63, Sox9, Actn1, Fzd7 and Krt17 were observed in our transgenic model. Our analysis presents an array of novel genes as putative corneal stem cell markers.

为了鉴定角膜上皮干细胞的可靠标志物，我们利用诱导型转基因“脉冲追踪”鼠模型（K5Tta×TRE-H2BGFP）来定位、纯化和鉴定角膜中的缓慢循环细胞。缓慢分裂细胞中 GFP 标记的保留允许将这些细胞定位到角膜缘，并通过 FACS 对其进行随后的纯化。与 GFP 快速分裂细胞相比，慢循环细胞中的转录组分析鉴定出差异表达的基因。将角膜上皮的 RNA-Seq 数据与表皮毛囊干细胞的 RNA-Seq 进行比较，以鉴定代表常见假定干细胞标志物或决定因素的基因，这些基因包括 Sox9、Fzd7、Actn1、Anxa3 和 Krt17。在我们的转基因模型中观察到 GFP 的保留和 Krt15、ΔNp63、Sox9、Actn1、Fzd7 和 Krt17 的免疫组织化学表达的重叠。我们的分析提出了一系列新的基因作为潜在的角膜干细胞标志物。