Isolation and enrichment of melanocytes from human corneal limbus using CD117 (c-Kit) as selection marker

使用 CD117 (c-Kit) 作为选择标记从人角膜缘分离和富集黑色素细胞

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作者:Naresh Polisetti, Ursula Schlötzer-Schrehardt, Thomas Reinhard, Günther Schlunck

Abstract

Limbal melanocytes (LM) are located in the basal epithelial layer of the corneoscleral limbus and interact with adjacent limbal epithelial progenitor cells. The exploration of their biological role in the maintenance of the limbal stem cell niche has been limited by the difficulty of LM isolation and cultivation. Here, we report on a facile protocol for the efficient isolation and enrichment of pure populations of human LMs by fluorescence-activated cell sorting (FACS) using antibodies raised against the cell surface marker CD117 (c-Kit). The enriched LMs retain self-renewal capacity and sustained melanin production, and are suitable to study the potential of LMs in stem cell-based corneal tissue engineering.

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