Characterization of corneal stromal stem cells with the potential for epithelial transdifferentiation.
Characterization of corneal stromal stem cells with the potential for epithelial transdifferentiation.
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DOI：
10.1186/scrt226
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发表时间：
2013-06-24
影响因子：
7.5
通讯作者：
Dua HS
中科院分区：
文献类型：
--
作者：
Hashmani K;Branch MJ;Sidney LE;Dhillon PS;Verma M;McIntosh OD;Hopkinson A;Dua HS
角膜基质越来越被认为是干细胞的储存库。与角膜缘和内皮小生境一样，基质干细胞通常位于周边角膜和利姆布斯中。已知这些外周和角膜缘基质细胞（PLCSC）在体外产生间充质干细胞。最近，一个共同的角膜基质和上皮祖细胞暗示。本研究旨在探讨角膜基质细胞的干细胞潜能及其上皮转分化能力。PLCSC在传统的基于Dulbecco改良Eagle培养基（DMEM）的角膜细胞培养基和基于M199的培养基中生长，并通过使用流式细胞术分析细胞表面标志物的分布，并分化成用定量聚合酶链反应（qPCR）和组织学染色分析的间充质表型。随后通过使用荧光激活细胞分选（FACS）基于⑶ 34和⑶ 105表达将M199中的PLCSC分成亚群。亚群的特征在于标记物和间充质分化能力。还培养完整的PLCSC和亚群用于上皮转分化。在M199中培养的细胞表现出更像干细胞的细胞表面标志物特征，并且角膜基质细胞标志物CD 34保留了几代，但在DMEM中培养的细胞中不存在。与DMEM相比，在M199中培养的细胞也表现出更大的间充质分化潜力。PLCSC可分为CD 34 + CD 105+、CD 34-CD 105+和CD 34-CD 105-亚群，其中CD 34 + CD 105+细胞在标志物表达和间充质分化潜力方面最具干细胞样。PLCSC亚群表现出不同的转分化成上皮表型的能力。最初为CD 34 + CD 105+的细胞显示出最大的分化潜力，产生CK 3+和CK 19+细胞，并表达一系列上皮祖细胞（HES 1、FRZB 1、DCT、SOD 2、ABCG 2、CDH 1、KRT 19）和终末分化（DSG 3、KRT 3、KRT 12、KRT 24）基因。培养液对PLCSCs的表型和分化能力有显著影响。间质含有异质细胞群，其中我们已将CD 34+细胞鉴定为具有间充质和上皮分化能力的干细胞群。
The corneal stroma is being increasingly recognized as a repository for stem cells. Like the limbal and endothelial niches, stromal stem cells often reside in the peripheral cornea and limbus. These peripheral and limbal corneal stromal cells (PLCSCs) are known to produce mesenchymal stem cells in vitro. Recently, a common corneal stromal and epithelial progenitor was hinted at. This study aims to examine the stem cell potential of corneal stromal cells and to investigate their epithelial transdifferentiation ability. PLCSCs were grown in traditional Dulbecco modified Eagle medium (DMEM)-based keratocyte culture medium and an M199-based medium and analyzed for a profile of cell-surface markers by using flow cytometry and differentiated into mesenchymal phenotypes analyzed with quantitative polymerase chain reaction (qPCR) and histologic staining. PLCSCs in M199 were subsequently divided into subpopulations based on CD34 and CD105 expression by using fluorescence- activated cell sorting (FACS). Subpopulations were characterized by marker profile and mesenchymal differentiation ability. Both whole PLCSCs and subpopulations were also cultured for epithelial transdifferentiation. Cells cultured in M199 demonstrated a more stem-like cell-surface marker profile, and the keratocyte marker CD34 was retained for several passages but absent in cells cultured in DMEM. Cells cultured in M199 also exhibited a greater mesenchymal differentiation potential, compared with DMEM. PLCSCs could be divided into CD34+CD105+, CD34-CD105+, and CD34-CD105- subpopulations, of which CD34+CD105+ cells were the most stemlike with regard to marker expression and mesenchymal differentiation potential. Subpopulations of PLCSCs exhibited differing abilities to transdifferentiate into epithelial phenotypes. Cells that were initially CD34+CD105+ showed the greatest differentiation potential, producing CK3+ and CK19+ cells, and expressed a range of both epithelial progenitor (HES1, FRZB1, DCT, SOD2, ABCG2, CDH1, KRT19) and terminally differentiated (DSG3, KRT3, KRT12, KRT24) genes. Culture medium has a significant effect on the phenotype and differentiation capacity of PLCSCs. The stroma contains a heterogeneous cell population in which we have identified CD34+ cells as a stem cell population with a capacity for mesenchymal and epithelial differentiation.
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影响因子：
29.4
作者：
Crosby, HA;Kelly, DA;Strain, AJ
通讯作者：
Strain, AJ
影响因子：
6.4
作者：
Elhalis, Hussain;Azizi, Behrooz;Jurkunas, Ula V.
通讯作者：
Jurkunas, Ula V.
影响因子：
3.9
作者：
Ahearne, M;Yang, Y;Liu, KK
通讯作者：
Liu, KK
影响因子：
3.4
作者：
Ahearne, Mark;Wilson, Samantha L.;Yang, Ying
通讯作者：
Yang, Ying
影响因子：
5.2
作者：
Du, Yiqin;Carlson, Eric C.;Funderburgh, Martha L.;Birk, David E.;Pearlman, Eric;Guo, Naxin;Kao, Winston W. -Y.;Funderburgh, James L.
通讯作者：
Funderburgh, James L.