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存在天然角膜缘基质细胞可提高体外培养角膜缘干细胞/祖细胞的扩增效率。

Presence of native limbal stromal cells increases the expansion efficiency of limbal stem/progenitor cells in culture.

机构信息

Cornea Division, Jules Stein Eye Institute, University of California, Los Angeles, 100 Stein Plaza, Los Angeles, CA 90095, USA.

出版信息

Exp Eye Res. 2013 Nov;116:169-76. doi: 10.1016/j.exer.2013.08.020. Epub 2013 Sep 7.

Abstract

Niche factors are important in the maintenance and regulation of stem cells. Limbal stromal cells are potentially a component of limbal stem cell (LSC) niche. We investigated the role of the limbal stromal cells in the ex vivo expansion of limbal stem/progenitor cells. Limbal epithelial cells were cultured as single-cell suspension and cell clusters from dispase II or collagenase A (ColA), or tissue explant. ColA isolated limbal stromal cells along with limbal epithelial cells. In the presence of limbal stromal cells, a higher absolute number of p63α(bright) cells (p < 0.05) and a higher proportion of K14 positive epithelial cells were obtained from both ColA and explant tissue cultures. Expansion of the stem/progenitor population from dispase isolation was more efficient in the form of cell clusters than single cell suspension based on the absolute number of p63α(bright) cells. Expansion of the stem cell population is similar in the single cell and cell cluster cultures that are derived from ColA isolation. Our finding suggests that limbal stromal cells and an intact cell-cell contact help to maintain LSCs in an undifferentiated state in vitro during expansion.

摘要

龛位因素在干细胞的维持和调控中起着重要作用。角膜缘基质细胞可能是角膜缘干细胞 (LSC) 龛位的一个组成部分。我们研究了角膜缘基质细胞在体外扩增角膜缘干细胞/祖细胞中的作用。将角膜缘上皮细胞作为单细胞悬液和用Dispase II 或胶原酶 A (ColA) 或组织外植体分离的细胞簇进行培养。ColA 分离出的角膜缘基质细胞与角膜缘上皮细胞一起。在角膜缘基质细胞存在的情况下,从 ColA 和组织外植体培养物中获得了更高数量的 p63α(bright)细胞(p<0.05)和更高比例的 K14 阳性上皮细胞。基于 p63α(bright)细胞的绝对数量,Dispase 分离的干细胞/祖细胞群体的扩增以细胞簇的形式比单细胞悬液更为有效。从 ColA 分离获得的单细胞和细胞簇培养物中,干细胞群体的扩增是相似的。我们的发现表明,在体外扩增过程中,角膜缘基质细胞和完整的细胞-细胞接触有助于维持 LSCs 的未分化状态。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/14ed/3900305/a93942083672/nihms523565f1.jpg

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