The Investigation of Hair Follicle Regeneration Utilizing Human Interfollicular Dermal and Epidermal Stem Cells

利用人毛囊间真皮和表皮干细胞进行毛囊再生的研究

基本信息

批准号：
17390474
负责人：
YOSHIMURA Kotaro
金额：
$ 10.05万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

项目摘要

Dermal papilla cells (DPCs) in the mammalian hair follicles have been shown to have an ability to induce and develop hair follicles. It is theoretically possible to regenerate human hair by expanding DPCs and transplanting them in the bald skin.We performed a microarray analysis and RT-PCR comparing human dermal papilla cells and dermal fibroblasts both obtained from adult scalp skin, in order to elucidate the genetic factors responsible for hair induction in tissue engineered hair follicles. In the microarray analysis we found that TGFs gene was significantly upregulated compared to dermal fibroblasts and that it was one of the candidate genes involving in hair regeneration. RT-PCR analysis revealed significant and sustained upregulation of TGFs mRNA in cultured dermal papilla cells compared to dermal fibroblasts. Furthermore, we found soluble factors secreted by keratinocytes significantly upregulated TGF beta-2 mRNA.Immunohistochemistry (IHC) of human scalp skin and hair follicles for the biomarkers previously reported as specific for IFKC was performed. The expression of the biomarkers in cultured keratinocytes was also investigated by FACS analysis and immunostaining. Keratin 15 (K15) was expressed in the bulge area in IHC. CD34 was expressed in the lower area, which includes the sub-bulge area and outer root sheath (ORS). CD271 (p75 neurotrophin receptor) was expressed in the lower ORS that was negative for K15. All of them were expressed only in the outermost layer adhering to the basement membrane.We examined reproducibility of two of in vivo models for trichogenesis previously reported, including 'chamber models', grafting cultured human DPCs into silicone chambers on nude mice, and 'sandwich models', implanting cultured human DPCs with rat sole skin under the skin of nude mice. 'Sandwich models' were more useful than 'chamber models' and other in vivo models for testing the inductive viability of human DPCs in terms of reproducibility.

哺乳动物毛囊中的真皮乳头细胞（DPC）已被证明具有诱导和发育毛囊的能力。理论上，通过扩增DPC并将其移植到秃头皮肤中可以再生人类头发。我们对从成人头皮皮肤中获得的人真皮乳头细胞和真皮成纤维细胞进行了微阵列分析和RT-PCR比较，以阐明遗传因素负责组织工程毛囊中的毛发诱导。在微阵列分析中，我们发现与真皮成纤维细胞相比，TGFs基因显着上调，并且它是参与毛发再生的候选基因之一。 RT-PCR 分析显示，与真皮成纤维细胞相比，培养的真皮乳头细胞中 TGFs mRNA 显着且持续上调。此外，我们发现角质形成细胞分泌的可溶性因子显着上调 TGF beta-2 mRNA。对人类头皮皮肤和毛囊进行了免疫组织化学 (IHC)，以检测先前报道为 IFKC 特异性的生物标志物。还通过 FACS 分析和免疫染色研究了培养的角质形成细胞中生物标志物的表达。 IHC 中角蛋白 15 (K15) 在凸起区域表达。 CD34在下部区域表达，包括亚隆起区和外根鞘（ORS）。 CD271（p75 神经营养素受体）在 K15 阴性的较低 ORS 中表达。所有这些都仅在粘附于基底膜的最外层中表达。我们检查了先前报道的两种毛发发生体内模型的重现性，包括“室模型”、将培养的人类 DPC 移植到裸鼠的硅胶室中和“三明治”模型中，将培养的人 DPC 植入裸鼠皮下的大鼠足底皮肤中。在测试人类 DPC 的诱导活力方面，“三明治模型”比“室模型”和其他体内模型在重现性方面更有用。