本文關鍵詞：毛囊干細胞的分離培養(yǎng)與鑒定　出處：《吉林大學》2009年碩士論文　論文類型：學位論文

  更多相關文章： 毛囊 毛囊干細胞 細胞培養(yǎng)

【摘要】： 創(chuàng)傷引起的皮膚組織缺損,傳統的治療方法是利用自體斷層皮片移植,但存在供皮量有限和造成額外損傷的障礙。利用體外培養(yǎng)的人工皮膚即組織工程皮膚作為移植的來源可以從根本上解決這個問題,但是如何獲得種子細胞卻一直困擾著人們。隨著近年來對干細胞研究的不斷深入,以干細胞作為種子細胞構建組織工程皮膚成為當前研究的熱點之一。 皮膚干細胞作為種子細胞構建組織工程皮膚是較好的選擇。皮膚干細胞主要有表皮干細胞和毛囊干細胞。目前,表皮干細胞作為種子細胞構建組織工程皮膚已經應用于臨床燒傷患者,但仍存在排異反應和供皮量有限等問題。 隨著醫(yī)學生物學的發(fā)展,皮膚組織工程的深入,毛囊干細胞作為一種新的種子細胞逐漸受到人們的關注。越來越多的研究表明毛囊中也存在具有多向分化潛能的干細胞,如果能建立完善的培養(yǎng)擴增體系,只需要一定數目的頭發(fā),即可培養(yǎng)出表皮的膜片,供給豐富,損傷小,不造成創(chuàng)面,無免疫排異反應,并可實現個體化治療,可為臨床皮膚缺損的修復開辟新的領域和提供更多的選擇方法。 有關毛囊干細胞分離培養(yǎng)的研究,目前在多數實驗中以鼠的觸須毛囊、外科獲取的人頭皮樣本為毛囊干細胞的組織來源。樣本經中性蛋白酶(dispaseⅡ)處理,分離去除表皮,從真皮中拔取并收集生長期的毛囊組織,進行毛囊干細胞組織塊培養(yǎng),或者用胰酶/EDTA處理收集的毛囊組織,獲得單細胞懸液培養(yǎng)毛囊干細胞。 本研究旨在從拔出的毛發(fā)中培養(yǎng)出毛囊干細胞,以求建立一種新的毛囊干細胞的獲取方法。實驗中,我們以直接拔取的人毛發(fā)作為組織來源,進行人毛囊干細胞原代培養(yǎng)方法的探索,并對其進行鑒定。通過比較幾種不同的毛囊干細胞原代培養(yǎng)方法,最終采用0.25%胰酶+0.05%EDTA消化毛囊組織,以絲裂霉素處理的人成纖維細胞為飼養(yǎng)層與毛囊組織及其來源的細胞共培養(yǎng),可以獲得毛囊組織來源的鋪路石樣細胞。經鑒定,此方法獲得的毛囊組織來源的鋪路石樣細胞中含有一定數量的毛囊干細胞。
[Abstract]:The traditional treatment for skin tissue defects caused by trauma is the use of autogenous skin grafts. However, there are limited skin donors and obstacles that cause additional damage. The use of tissue engineered skin as a source of transplantation can fundamentally solve this problem. However, how to obtain seed cells has been puzzling people. With the development of stem cell research in recent years, using stem cells as seed cells to construct tissue engineered skin has become one of the hot research topics. Skin stem cells as seed cells to construct tissue engineered skin is a good choice. Skin stem cells are mainly epidermal stem cells and hair follicle stem cells. Epidermal stem cells as seed cells for tissue engineering skin have been used in clinical burn patients, but there are still some problems such as rejection and limited skin supply. With the development of medical biology, skin tissue engineering is becoming more and more important. As a new seed cell, hair follicle stem cells have attracted more and more attention. More and more researches show that there are stem cells with multi-differentiation potential in hair follicles, if a perfect culture and amplification system can be established. Only a certain number of hair can be cultured out of the epidermis membrane, supply rich, small damage, no wound, no immune rejection reaction, and can achieve individualized treatment. It can open up a new field and provide more options for the repair of clinical skin defects. Hair follicle stem cells have been isolated and cultured in most experiments with mouse tentacles. The human scalp samples obtained by surgery were the tissue source of hair follicle stem cells. The samples were treated with neutral protease dispase 鈪，

本文編號：1382440