Characterization and isolation of stem cell-enriched human hair follicle bulge cells
Characterization and isolation of stem cell-enriched human hair follicle bulge cells
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DOI：
10.1172/jci26043
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发表时间：
2006-01-01
影响因子：
15.9
通讯作者：
Vogel, JC
中科院分区：
文献类型：
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作者：
Ohyama, M;Terunuma, A;Vogel, JC
人毛囊隆突是角质形成干细胞（KSC）的重要生态位。通过分析全球基因表达谱和鉴定独特的细胞表面标记物可以促进人类隆突细胞生物学的阐明。人类毛囊缺乏独特的隆突形态，阻碍了隆突细胞和KSC的研究。在这项研究中，我们确定了标记保留细胞的分布，以确定人类生长期隆起。利用导航激光捕获显微切割技术，从其他卵泡区域获得隆突细胞和外根鞘细胞，并利用cDNA微阵列进行分析。基因转录物编码抑制剂的WNT和激活素/骨形态发生蛋白信号被过度的隆起，而负责细胞增殖的基因被低估，与生长期卵泡中存在静止的非周期性KSC一致。隆突细胞的阳性标志物包括CD 200、PHLDA 1、卵泡抑素和卷曲同源物1，而CD 24、CD 34、CD 71和CD 146优先由非隆突角质形成细胞表达。重要的是，从毛囊悬浮液中获得的CD 200（+）细胞（CD 200（hi）CD 24（lo）CD 34（lo）CD 71（lo）CD 146（lo））在克隆形成试验中显示出高集落形成效率，表明成功富集了活的人隆突干细胞。富集隆突细胞的干细胞行为及其在基因治疗和毛发再生中的效用需要在体内测定中进行评估。
The human hair follicle bulge is an important niche for keratinocyte stem cells (KSCs). Elucidation of human bulge cell biology could be facilitated by analysis of global gene expression profiles and identification of unique cell-surface markers. The lack of distinctive bulge morphology in human hair follicles has hampered studies of bulge cells and KSCs. In this study, we determined the distribution of label-retaining cells to define the human anagen bulge. Using navigated laser capture microdissection, bulge cells and outer root sheath cells from other follicle regions were obtained and analyzed with cDNA microarrays. Gene transcripts encoding inhibitors of WNT and activin/bone morphogenic protein signaling were overrepresented in the bulge, while genes responsible for cell proliferation were underrepresented, consistent with the existence of quiescent noncycling KSCs in anagen follicles. Positive markers for bulge cells included CD200, PHLDA1, follistatin, and frizzled homolog 1, while CD24, CD34, CD71, and CD146 were preferentially expressed by nonbulge keratinocytes. Importantly, CD200(+) cells (CD200(hi)CD24(lo)CD34(lo)CD71(lo)CD146(lo)) obtained from hair follicle suspensions demonstrated high colony-forming efficiency in clonogenic assays, indicating successful enrichment of living human bulge stem cells. The stem cell behavior of enriched bulge cells and their utility for gene therapy and hair regeneration will need to be assessed in in vivo assays.