Department of Biology, Faculty of Basic Sciences, University of Shahrekord, Shahrekord, Iran.
In Vitro Cell Dev Biol Anim. 2010 Jan;46(1):54-9. doi: 10.1007/s11626-009-9245-y.
In healthy individuals, skin integrity is maintained by epidermal stem cells which self-renew and generate daughter cells that undergo terminal differentiation. Epidermal stem cells represent a promising source of stem cells, and their culture has great potential in scientific research and clinical application. However, no single method has been universally adopted for identifying and isolating epidermal stem cells. Here, we reported the isolation and characterization of putative epidermal stem cells from newborn mouse skin. The keratinocytes were separated enzymatically. Putative epidermal stem cells were selected by rapid adherence on a composite matrix made of type I collagen and fibronectin. Unattached cells were discarded after 10 min, and the attached cells were cultured in a defined culture medium. The isolated cells showed the typical epidermal stem cell morphology. Immunofluorescence indicated that the cells were strongly stained for β1 integrin family of extracellular matrix receptors. In conclusion, mouse putative epidermal stem cells were successfully isolated from newborn mouse epidermis on the basis of high rapid adhesion to extracellular matrix proteins and cultured in vitro.
在健康个体中，表皮干细胞通过自我更新并产生经历终末分化的子细胞来维持皮肤完整性。表皮干细胞是一种很有前途的干细胞来源，其培养在科学研究和临床应用中有巨大的潜力。然而，目前还没有一种普遍适用的方法来鉴定和分离表皮干细胞。在这里，我们报道了从新生小鼠皮肤中分离和鉴定表皮干细胞的方法。通过酶消化分离角蛋白细胞。将角蛋白细胞接种于由Ⅰ型胶原和纤维连接蛋白组成的复合基质上，通过快速黏附来选择表皮干细胞。10 分钟后去除未黏附的细胞，将黏附的细胞在限定条件的培养基中培养。分离得到的细胞表现出典型的表皮干细胞形态。免疫荧光染色表明，细胞强烈表达β1 整合素家族细胞外基质受体。总之，我们成功地基于细胞对细胞外基质蛋白的高快速黏附特性从新生鼠表皮中分离出表皮干细胞，并在体外进行培养。
