骨髓间充质干细胞体外诱导分化为表皮干细胞的研究

发布时间：2018-05-02 10:07

本文选题：骨髓间充质干细胞 + 表皮干细胞　；参考：《中南大学》2006年博士论文

【摘要】：背景：烧伤、创伤和整形外科的创面修复需要大量自体皮肤，组织工程化皮肤是解决自体皮来源的良好方法。理想的组织工程化皮肤应该是能在尽量短的时间内合成包含有表皮、真皮和各种皮肤附件(汗腺、毛囊、皮脂腺等)以及黑色素细胞和朗格罕氏细胞的永久性覆盖的皮肤，但至今还没有任何一种组织工程皮肤能作为理想的皮肤覆盖物，其中种子细胞和支架材料是组织工程研究的关键。表皮干细胞具有多分化潜能，除再生皮肤的多层表皮结构外，还形成汗腺、毛发和毛囊等皮肤附属器官，但由于目前获取与纯化表皮干细胞较为困难，从而妨碍了表皮干细胞作为组织工程化皮肤种子细胞的应用。本研究以HaCaT细胞培养上清液联合EGF作为诱导条件，观测BMSCs经诱导后的细胞形态学及细胞生物化学的变化，以明确BMSCs能否在体外诱导分化为表皮干细胞；通过以BrdU标记BMSCs后，植入全层皮肤缺损创面，观察其在皮肤缺损创面愈合中的作用，了解骨髓间充质干细胞能否在体内诱导分化表皮干细胞；并观察其在明胶／聚己酸内酯电纺纳米支架中生长的情况。为选用骨髓间充质干细胞作为组织工程化皮肤的种子细胞提供理论依据。第一章：兔自体骨髓间充质干细胞促进创面愈合的实验研究目的：观察骨髓间充质干细胞(Bone Mesenchymal Stem Cells，BMSCs)在皮肤缺损创面愈合中的作用，，探讨其作为种子细胞修复与重建皮肤的可能性。方法：取成年家兔30只，抽取骨髓，采用直接贴壁法分离、纯化和培养骨髓间充质干细胞；采用原位杂交试剂盒对培养细胞进行cDNA探针原位杂交检测CD44，鉴定其为骨髓间充质干细胞；于兔背部两侧制造全层皮肤缺损创面，以Ⅰ型胶原为载体，将已用5-溴脱氧尿嘧啶(5-bromodeoxy-uridine，BrdU)标记的自体BMSCs植于左侧创面，
[Abstract]:Background: the repair of burn, trauma and orthopedic wounds requires a large number of autologous skin. Tissue engineered skin is a good method to solve the source of autogenous skin. Ideal tissue engineered skin should be permanently covered with epidermis, dermis and various skin attachments (sweat glands, hair follicles, sebaceous glands, etc.) and melanocytes and Langerhans cells in as short a time as possible. However, no tissue engineered skin can be used as an ideal skin covering, and seed cells and scaffolds are the key to tissue engineering. Epidermal stem cells have the potential of multi-differentiation. Besides the multi-layer epidermal structure of regenerated skin, epidermal stem cells also form sweat glands, hair and hair follicles. However, it is difficult to obtain and purify epidermal stem cells. This hinders the application of epidermal stem cells as seed cells of tissue engineering skin. In this study, the supernatant of HaCaT cell culture combined with EGF was used as the induction condition to observe the changes of cell morphology and cell biochemistry of BMSCs after induction, to determine whether BMSCs could be induced to differentiate into epidermal stem cells in vitro, and to determine whether BMSCs could be induced into epidermal stem cells by BrdU labeled BMSCs. To investigate whether the bone marrow mesenchymal stem cells can induce the differentiation of epidermal stem cells in vivo, the effect of the whole skin defect wound was observed. Its growth in gelatin / polycaprolactone electrospun nano-scaffold was observed. To provide theoretical basis for selecting bone marrow mesenchymal stem cells as seed cells of tissue engineered skin. Chapter 1: experimental study on promoting wound healing by autologous bone marrow mesenchymal stem cells in rabbits Objective: to observe the role of bone Mesenchymal Stem cells (BMSCs) in the healing of skin defect wounds, and to explore the possibility of repairing and reconstructing the skin as seed cells of bone marrow mesenchymal stem cells (BMSCs). Methods: bone marrow was extracted from 30 adult rabbits. Bone marrow mesenchymal stem cells were purified and cultured by direct adherent method. CD44 was detected by in situ hybridization with cDNA probe and identified as bone marrow mesenchymal stem cells by in situ hybridization kit. Autologous BMSCs labeled with 5-bromodeoxy-uridine (BrdU) was implanted on the left wound.
【学位授予单位】：中南大学
【学位级别】：博士
【学位授予年份】：2006
【分类号】：R329.2

【参考文献】