人角膜缘上皮祖细胞体外杯状细胞分化潜能。

Goblet Cell Differentiation Potential in Human Corneal Limbal Epithelial Progenitor Cells In Vitro.

机构信息

Department of Ophthalmology, University of Tokyo Graduate School of Medicine, Tokyo, Japan.

Department of Ophthalmology, Kyoto Prefectural University of Medicine, Japan.

出版信息

Invest Ophthalmol Vis Sci. 2020 Oct 1;61(12):27. doi: 10.1167/iovs.61.12.27.

DOI:10.1167/iovs.61.12.27

PMID:33112944

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7594597/

Abstract

PURPOSE

The existence of goblet cells has been regarded as a critical differential point to distinguish conjunctival epithelium from corneal epithelium in vivo. We tested differentiation potential of single progenitor cells from corneal limbal epithelium with growth factors in vitro.

METHODS

Dissociated single cells from corneal limbal epithelium were cultured in the serum- and feeder cell-free medium containing B27 and various growth factors using nontissue culture dishes. Specific marker expression was examined in the colonies stimulated with growth factors. Differentiation of some mucosal epithelia was tested.

RESULTS

Adherent single cells from dissociated single cells in corneal limbal epithelium did not proliferate in the serum- and feeder cell-free medium containing B27 only and formed corneal epithelium with B27 plus epidermal growth factor, while they gave rise to goblet cell with periodic acid Schiff-positive mucin and cytokeratin-3 and-12 expressing corneal epithelium with fibroblast growth factor (FGF)2 stimulation. Colonies stimulated with FGF2 expressed goblet cell specific MUC5AC and cytokeratin-7 mRNA and protein. FGF receptor 1 was a functional receptor for the differentiation to goblet cells and corneal epithelium.

CONCLUSIONS

Single corneal limbal progenitor cells give rise to goblet cells and corneal epithelium by FGF2 stimulation via FGF receptor 1 in vitro.

摘要

目的

杯状细胞的存在被认为是区分活体结膜上皮和角膜上皮的一个关键鉴别点。我们在体外利用生长因子测试了角膜缘上皮的单个前体细胞的分化潜能。

方法

使用无组织培养皿，在含有 B27 和各种生长因子的无血清和无饲养细胞的培养基中培养角膜缘上皮分离的单个细胞。在生长因子刺激下检测特定标志物的表达情况。测试了一些黏膜上皮的分化情况。

结果

角膜缘上皮分离的单个细胞的贴壁单细胞在仅含有 B27 的无血清和无饲养细胞的培养基中不增殖，用 B27 和表皮生长因子形成角膜上皮，而用成纤维细胞生长因子（FGF）2 刺激则形成具有帕氏阳性粘蛋白和角蛋白-3 和-12 表达的杯状细胞的角膜上皮。用 FGF2 刺激的集落表达杯状细胞特异性 MUC5AC 和角蛋白-7mRNA 和蛋白。FGF 受体 1 是向杯状细胞和角膜上皮分化的功能性受体。