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.

哺乳动物毛囊中的皮肤乳头细胞（DPC）已被证明具有诱导和发展毛囊的能力。从理论上讲，可以通过扩展DPC并在秃头皮肤中移植人头发来再生。在微阵列分析中，我们发现与皮肤成纤维细胞相比，TGFS基因显着上调，并且它是涉及毛发再生的候选基因之一。 RT-PCR分析显示，与皮肤成纤维细胞相比，培养的皮肤乳头细胞中TGFS mRNA的显着和持续上调。此外，我们发现，角质形成细胞分泌的可溶性因子显着上调了人头皮皮肤和毛囊的免疫组织化学（IHC）先前报道为IFKC的特异性生物标志物。还通过FACS分析和免疫染色研究了生物标志物在培养的角质形成细胞中的表达。角蛋白15（K15）在IHC的凸出区域表示。 CD34在较低区域表达，其中包括子块区域和外部鞘（OR）。 CD271（p75神经营养受体）在K15阴性的下OR中表达。所有这些都仅在粘附在地下膜上的最外层中表达。我们检查了先前报道的两个体内模型的可重复性，包括“室模型”，将培养的人类DPC接管裸小鼠的硅胶腔，以及植入培养的人类DPC的硅质腔，并植入了与人dpc的植入式人DPC，含有bat s bat in Cat in Cat in Cat in Cat nefer Rat s ncter Sprice nude nude nude nude nude nude nude nude nude nude nude nude nude nude nude nude nude nude nude nude nude nude nude nude。 “三明治模型”比“室模型”和其他体内模型更有用，该模型在可重复性方面测试了人类DPC的感应性。