Program in Tissue Engineering, Department of Parasitology, Nara Medical University, 840 Shijo-cho, Kashihara, Nara 634-8521, Japan.
J Biosci Bioeng. 2010 Aug;110(2):217-22. doi: 10.1016/j.jbiosc.2010.01.020. Epub 2010 Mar 3.
Although mouse Wnt-10b has been shown to play various roles in a wide range of biological actions, the effects on epithelial stem/progenitor cells in the skin have not been reported. In the present study, we investigated the effects of Wnt-10b on proliferation and differentiation of murine skin-derived CD34 and CD49f double-positive (CD34(+)CD49f(+)) cells, a supposed fraction as enriched epithelial stem/progenitor cells. The cells were prepared from dorsal skin samples obtained from young adult mice as alpha6 integrin (CD49f) and CD34 double-positive cells by fluorescent activated cell sorting (FACS), and they were cultured with or without Wnt-10b to investigate its effects on proliferation and differentiation. Involvement of canonical Wnt signaling pathway was confirmed by TOPFLASH assay, and differentiation of the CD34(+)CD49f(+) cells was assessed by RT-PCR analysis and immunocytochemical examinations. The skin-derived CD34(+)CD49f(+) cells were immunopositive for Lhx2 and expressed mRNA of classical markers for bulge stem cells, including Lhx2, keratin15, Sox9, S100a6, and NFATc1. Their proliferation was suppressed by Wnt-10b, and the markers for differentiated epithelial cells became to be expressed in the culture with Wnt-10b. These results suggest that Wnt-10b promotes differentiation of epithelial stem/progenitor cells in the skin.
尽管已经证明小鼠 Wnt-10b 在广泛的生物学作用中发挥了各种作用，但它对皮肤上皮干细胞/祖细胞的影响尚未报道。在本研究中，我们研究了 Wnt-10b 对小鼠皮肤来源的 CD34 和 CD49f 双阳性（CD34(+)CD49f(+)）细胞增殖和分化的影响，CD34(+)CD49f(+)细胞被认为是富含上皮干细胞/祖细胞的细胞。这些细胞是从小鼠背部皮肤样本中通过荧光激活细胞分选（FACS）获得的 α6 整合素（CD49f）和 CD34 双阳性细胞，然后在有或没有 Wnt-10b 的情况下培养，以研究其对增殖和分化的影响。通过 TOPFLASH 测定证实了经典 Wnt 信号通路的参与，通过 RT-PCR 分析和免疫细胞化学检查评估 CD34(+)CD49f(+)细胞的分化。皮肤来源的 CD34(+)CD49f(+)细胞对 Lhx2 呈免疫阳性反应，并表达经典毛球干细胞标志物的 mRNA，包括 Lhx2、角蛋白 15、Sox9、S100a6 和 NFATc1。Wnt-10b 抑制它们的增殖，并且在有 Wnt-10b 的培养中，分化的上皮细胞标志物开始表达。这些结果表明 Wnt-10b 促进皮肤上皮干细胞/祖细胞的分化。
