Inoue Keita, Aoi Noriyuki, Sato Takahiro, Yamauchi Yuji, Suga Hirotaka, Eto Hitomi, Kato Harunosuke, Araki Jun, Yoshimura Kotaro
Department of Plastic Surgery, University of Tokyo, Tokyo, Japan.
Lab Invest. 2009 Aug;89(8):844-56. doi: 10.1038/labinvest.2009.48. Epub 2009 Jun 8.
Several putative biomarkers have been suggested for identifying murine follicular stem cells; however, human hair follicles have a different pattern of biomarker expression, and follicular stem cell isolation methods have not been established. To isolate a stem cell population applicable to clinical settings, we conducted a comprehensive survey of the expression of stem-cell-associated (K15, CD200, CD34, and CD271) and other biomarkers (K1, K14, CD29, and CD49f) in immunohistological sections of the human epidermis and follicular outer root sheath (ORS). We also examined freshly isolated and cultured epidermal or follicular cells with single- and multicolor flow cytometry or immunocytochemistry. After sorting cells by CD200, CD34, and forward scatter (FSC) values (cell size), colony-forming assays were performed. We found that biomarkers were differentially expressed in the epidermis and ORS. Basal bulge cells were mainly K15+CD200+CD34(-)CD271(-), and suprabasal cells were K15(-)CD200+CD34(-)CD271(-). We categorized follicular cells into nine subpopulations according to biomarker expression profiles. The CD200+CD34(-) bulge cells had much higher colony-forming abilities than the CD34+ population, and were divided into two subpopulations: a CD200+CD34(-)FSC(high) (K15-rich, basal) and a CD200+CD34(-)FSC(low) (K15-poor, suprabasal) population. The former formed fewer but larger-sized colonies than the latter. Follicular epithelial cell cultivation resulted in loss of K15, CD200, CD34, and CD271 expression, but maintenance of K14, CD29, and CD49f expression. We found that the bulge contained two populations with different localizations, cell sizes, and colony-forming abilities. We showed that K15, CD200, CD34, and CD271 were useful biomarkers for characterizing freshly isolated human follicular epithelial cells in diverse stages of differentiation.
已经提出了几种假定的生物标志物用于鉴定小鼠毛囊干细胞；然而，人类毛囊具有不同的生物标志物表达模式，并且毛囊干细胞分离方法尚未建立。为了分离适用于临床的干细胞群体，我们对人类表皮和毛囊外根鞘（ORS）免疫组织学切片中干细胞相关标志物（K15、CD200、CD34和CD271）以及其他生物标志物（K1、K14、CD29和CD49f）的表达进行了全面调查。我们还通过单颜色和多颜色流式细胞术或免疫细胞化学检查了新鲜分离和培养的表皮或毛囊细胞。通过CD200、CD34和前向散射（FSC）值（细胞大小）对细胞进行分选后，进行了集落形成试验。我们发现生物标志物在表皮和ORS中差异表达。基底隆突细胞主要为K15+CD200+CD34(-)CD271(-)，而上基底细胞为K15(-)CD200+CD34(-)CD271(-)。我们根据生物标志物表达谱将毛囊细胞分为九个亚群。CD200+CD34(-)隆突细胞的集落形成能力比CD34+群体高得多，并分为两个亚群：一个CD200+CD34(-)FSC(高)（富含K15，基底）群体和一个CD200+CD34(-)FSC(低)（K15贫乏，上基底）群体。前者形成的集落比后者少但更大。毛囊上皮细胞培养导致K15、CD200、CD34和CD271表达丧失，但K14、CD29和CD49f表达得以维持。我们发现隆突包含两个具有不同定位、细胞大小和集落形成能力的群体。我们表明，K15、CD200、CD34和CD271是用于表征不同分化阶段新鲜分离的人类毛囊上皮细胞的有用生物标志物。
