以角蛋白水膠培養真皮乳頭細胞

Culturing dermal papilla cells on keratin-based hydrogels

劉宇晨(Yu-Chen Liu)

指導教授 ： 黃義侑

國立臺灣大學/工學院/醫學工程學研究所/碩士(2017年)

https://doi.org/10.6342/NTU201703653

摘要

禿髮為一種常見的病症，雖然禿髮不會威脅到生命，但對病人的社交活動以及心理健康都會造成深遠的影響。其中雄性禿是造成落髮最常見的原因，由於遺傳基因以及雄性荷爾蒙的影響，造成毛囊萎縮。現行治療方法包含藥物治療或者是植髮。目前美國食品藥物管理局認可的療法包含口服的藥物finasteride以及外用的生髮水minoxidil。在毛囊重建研究中，真皮乳頭細胞是最早被探討以及大量研究的，現已有大量的研究證實真皮乳頭細胞不論在體內或體外培養，皆有誘導毛囊再生的能力。然而隨著繼代培養後，其誘導能力會漸漸消失。
頭髮的角蛋白是一種細胞骨架蛋白，在頭髮生成的周期中，由毛囊內的表皮細胞在頭髮的生長期中生成角蛋白為基礎的材料被應用在生醫領域的包括周邊神經的再生、傷口敷料、止血劑，角膜的重建以及藥物釋放的載體等等。本研究旨在探討利用角蛋白所構成之水膠以及薄膜是否能使真皮乳頭細胞在高代的培養下還能維持其誘導能力。
利用氧化法及還原法能成功的從頭髮萃取出角蛋白keratose 及kerateine，並利用genipin為交聯劑，成功製備出水膠以及薄膜的型態。實驗結果顯示，真皮乳頭細胞在角蛋白材料上有別於培養皿培養的培養狀態，並利用培養皿一半未修飾、一半披覆角蛋白水膠確認此結果是由材料與細胞之間的交互作用所造成的，透過免疫螢光的染色，確認在其上培養能保有其誘導能力。真皮乳頭細胞在keratose水膠上的幹細胞特性遠遠大於在培養皿中培養，或是在明膠水膠上培養。應用此材料於未來體外大量培養頭髮的真皮乳頭細胞應該很有發展的潛力。

關鍵字

角蛋白 ； 水膠 ； 真皮乳頭細胞 ； 交聯劑 ； 細胞成團

並列摘要

Alopecia is a common disease that would make huge impact on patients’ mental health and social life. Among all the causes, androgenetic alopecia is the most common cause of alopecia which caused by genetic issue and excessive androgen, resulting in the atrophy of hair follicle. Current treatment includes medicine or hair transplant. The only two US Food and Drug Administration certified therapies include oral administration of finasteride as well as topical hair water minoxidil. In the study of regeneration of hair follicle, dermal papilla were first explored and extensively studied.. It is proved that both in vitro and in vivo culturing have the ability to induce hair follicle regeneration. Unfortunately. After subculture, the ability disappear.
Hair keratin is a kind of cytoskeleton that be made during the hair cycle. It is made by the epithelial cell in the hair follicle in anangen. The keratin based material can be used in the biomedical research, including the regeneration of periphery nerve system, wound healing, hemostat, cornea restruction and drug delivery system.
This study aims to investigate whether keratin-based material could maintain the ability of inducing hair follicle regeneration of dermal papilla.
We successfully extracted keratose and kerateine by oxidization and reduction method. Using genipin as cross linker, we were able to make film and hydrogel based on keratin. According to the results, dermal papilla cells show different morphology on keratose hydrogel when comparing with normal culture plate. By coating half keratose film on the culture plate, we confirm that the different result of culturing dermal papilla cells was caused by the interaction between the cells and the materials. By immmunostaining, we confirm that culturing cells on keratose hydrogel were able to maintain its ability to induce hair follicle regeneration. Culturing dermal papilla cells on keratose hydrogel could maintain its stemness over culturing on normal tissue plate or on gelatin hydrogel. The material has potential for culturing large quantity dermal papilla cells in vitro.

並列關鍵字

keratin ； hydrogel ； dermal papilla ； cross linker ； cell sphere

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