干细胞
表皮(动物学)
细胞生物学
人口
生物
成体干细胞
碘化丙啶
边居
祖细胞
单元格排序
癌症干细胞
分子生物学
细胞分化
流式细胞术
解剖
生物化学
基因
细胞凋亡
医学
环境卫生
程序性细胞死亡
作者
Martine Dunnwald,Ann Tomanek‐Chalkley,Dana Alexandrunas,Justin Fishbaugh,Jackie R. Bickenbach
标识
DOI:10.1034/j.1600-0625.2001.100106.x
摘要
Abstract: Continuously renewing tissues, such as the epidermis, are maintained by stem cells that slowly proliferate and remain in the tissue for life. Although it has been known for decades that epithelial stem cells can be identified as label‐retaining cells (LRCs) by long term retention of a nuclear label, isolating a pure population of stem cells has been problematic. Using a Hoechst and propidium iodide dye combination and specifically defined gating, we sorted mouse epidermal basal cells into three fractions, which we have now identified as stem, transient amplifying (TA), and nonproliferative basal cells. More than 90% of freshly isolated stem cells showed a G 0 /G 1 cell cycle profile, while greater than 20% of the TA cells were actively dividing. Both stem and TA cells retained proliferative capacity, but the stem cells formed larger, more expandable colonies in culture. Both populations could be transduced with a retroviral vector and used to bioengineer an epidermis. However, only the epidermis from the stem cell population continued to grow and express the reporter gene for 6 months in organotypic culture. The epidermis from the transient amplifying cell fraction completely differentiated by 2 months. This novel sorting method yields pure viable epithelial stem cells that can be used to bioengineer a tissue and to test permanent recombinant gene expression.
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