毛囊干细胞的分离培养与鉴定

发布时间：2018-01-05 09:06

  本文关键词：毛囊干细胞的分离培养与鉴定　出处：《吉林大学》2009年硕士论文　论文类型：学位论文

  更多相关文章： 毛囊 毛囊干细胞 细胞培养

【摘要】： 创伤引起的皮肤组织缺损,传统的治疗方法是利用自体断层皮片移植,但存在供皮量有限和造成额外损伤的障碍。利用体外培养的人工皮肤即组织工程皮肤作为移植的来源可以从根本上解决这个问题,但是如何获得种子细胞却一直困扰着人们。随着近年来对干细胞研究的不断深入,以干细胞作为种子细胞构建组织工程皮肤成为当前研究的热点之一。 皮肤干细胞作为种子细胞构建组织工程皮肤是较好的选择。皮肤干细胞主要有表皮干细胞和毛囊干细胞。目前,表皮干细胞作为种子细胞构建组织工程皮肤已经应用于临床烧伤患者,但仍存在排异反应和供皮量有限等问题。 随着医学生物学的发展,皮肤组织工程的深入,毛囊干细胞作为一种新的种子细胞逐渐受到人们的关注。越来越多的研究表明毛囊中也存在具有多向分化潜能的干细胞,如果能建立完善的培养扩增体系,只需要一定数目的头发,即可培养出表皮的膜片,供给丰富,损伤小,不造成创面,无免疫排异反应,并可实现个体化治疗,可为临床皮肤缺损的修复开辟新的领域和提供更多的选择方法。 有关毛囊干细胞分离培养的研究,目前在多数实验中以鼠的触须毛囊、外科获取的人头皮样本为毛囊干细胞的组织来源。样本经中性蛋白酶(dispaseⅡ)处理,分离去除表皮,从真皮中拔取并收集生长期的毛囊组织,进行毛囊干细胞组织块培养,或者用胰酶/EDTA处理收集的毛囊组织,获得单细胞悬液培养毛囊干细胞。 本研究旨在从拔出的毛发中培养出毛囊干细胞,以求建立一种新的毛囊干细胞的获取方法。实验中,我们以直接拔取的人毛发作为组织来源,进行人毛囊干细胞原代培养方法的探索,并对其进行鉴定。通过比较几种不同的毛囊干细胞原代培养方法,最终采用0.25%胰酶+0.05%EDTA消化毛囊组织,以丝裂霉素处理的人成纤维细胞为饲养层与毛囊组织及其来源的细胞共培养,可以获得毛囊组织来源的铺路石样细胞。经鉴定,此方法获得的毛囊组织来源的铺路石样细胞中含有一定数量的毛囊干细胞。
[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，

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