本文選題：外根鞘細(xì)胞　切入點：毛乳頭細(xì)胞　出處：《昆明醫(yī)學(xué)院》2010年碩士論文　論文類型：學(xué)位論文

【摘要】： 目的:自行分離培養(yǎng)獲得和購買ScienCell公司的人毛囊外根鞘細(xì)胞株(HHFORSC)及人毛囊毛乳頭細(xì)胞株(HHDPC),以得到充足的種子細(xì)胞。HHDPC、HHFORSC與海藻酸鈉3D支架在體外構(gòu)建三維模型,培養(yǎng)后植入SD大鼠皮下以誘導(dǎo)其形成毛囊,并探討毛囊的生物學(xué)特征和這兩種細(xì)胞在毛囊形成中的作用。 方法:購買ScienCell公司的人毛囊外根鞘細(xì)胞株(HHFORSC)和人毛囊毛乳頭細(xì)胞株(HHDPC),復(fù)蘇培養(yǎng);同時利用酶消化法和組織塊法獲得HHDPC和HHFORSC,培養(yǎng)后比較兩種不同途徑得到的細(xì)胞生長特征,以獲得充足的HHFORSC和HHDPC。將上述兩種細(xì)胞混合后種植于海藻酸鈉3D細(xì)胞支架,構(gòu)建毛囊的體外三維模型,培養(yǎng)7周,光鏡觀察毛囊形成情況。同時將該三維模型移植入SD大鼠皮下,飼養(yǎng)SD大鼠8周,移植部位取材后,行HE染色、免疫組化和電鏡觀察毛囊形成情況。 結(jié)果:由ScienCell公司購買的細(xì)胞株和我們自行分離培養(yǎng)獲得的HHDPC和HHFORSC生長特征沒有明顯差異。體外毛囊三維模型培養(yǎng)7周后,光鏡下可見支架中有均勻散在分布的毛囊混合細(xì)胞,未見到角化物質(zhì)及毛囊結(jié)構(gòu)。SD大鼠飼養(yǎng)8周移植部位取材,HE染色可見細(xì)胞聚集成團(tuán)狀的毛囊樣結(jié)構(gòu),CK14,CK15和波形蛋白染色陽性,電鏡下可見支架中有散在細(xì)胞分布。 結(jié)論:我們通過酶消化法和組織塊法分離獲得的HHDPC和HHFORSC保持了良好的生長特性。體外毛囊三維模型培養(yǎng)未能形成毛囊樣結(jié)構(gòu)。SD大鼠體內(nèi)三維模型培養(yǎng),HE染色可見細(xì)胞聚集成毛囊樣結(jié)構(gòu),CK14,CK15染色陽性,波形蛋白染色陽性,證明在該模型中誘導(dǎo)出了具有向毛囊分化傾向的毛囊樣結(jié)構(gòu),并且HHDPC保持了誘導(dǎo)毛囊形成的能力,HHFORSC保持了毛囊干細(xì)胞的作用,為以后成功重建毛囊指明了方向。
[Abstract]:Objective: to isolate and culture the human hair follicle outer root sheath cell line HHFORSCand the human hair follicle hair papilla cell line HHDPCC, so as to obtain sufficient seed cells, HHDPC-HHFORSC and sodium alginate 3D scaffolds to construct a three-dimensional model in vitro. After culture, SD rats were implanted subcutaneously to induce the formation of hair follicles. The biological characteristics of hair follicles and the role of these two kinds of cells in hair follicle formation were discussed. Methods: human hair follicle outer root sheath cell line HHFORSC) and human hair follicle dermal papilla cell line HHDPCC were purchased for resuscitation culture, and HHDPC and HHFORSCwere obtained by enzyme digestion and tissue block method. In order to obtain sufficient HHFORSC and HHDPC. the above two kinds of cells were mixed and implanted in sodium alginate 3D cell scaffold. The three-dimensional model of hair follicle was established in vitro and cultured for 7 weeks. The formation of hair follicle was observed by light microscope. At the same time, the model was transplanted subcutaneously into SD rats. SD rats were fed for 8 weeks. After transplantation, he staining, immunohistochemistry and electron microscope were used to observe the hair follicle formation. Results: there was no significant difference between the growth characteristics of HHDPC and HHFORSC obtained by ScienCell and the growth characteristics of HHDPC and HHFORSC obtained by our own isolation and culture. After 7 weeks of culture in vitro hair follicle, there were hair follicle mixed cells scattered in the scaffold under light microscope. No keratocystis and hair follicles were observed in SD rats after 8 weeks of feeding. He staining showed that the cells were clustered into clusters of hair follicle structure, CK14, CK15 and vimentin were positive, and the cells were scattered in the scaffold under electron microscope. Conclusion: the HHDPC and HHFORSC isolated by enzyme digestion and tissue mass method have good growth characteristics. In vitro hair follicle three-dimensional model culture can not form hair follicle-like structure. SD rat three-dimensional model culture can be stained with HE. It was found that the cells gathered to form a follicle-like structure, and the cells were positive for CK14 / CK-15 staining. The positive staining of vimentin showed that hair follicle-like structure was induced in the model, and HHDPC maintained the ability to induce hair follicle formation. HHFORSC maintained the role of hair follicle stem cells. It points out the direction for the successful reconstruction of hair follicles in the future.
【學(xué)位授予單位】：昆明醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R751

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