GAP-43在骨髓间充质干细胞向神经样细胞分化过程中作用机制研究

发布时间：2018-03-30 23:20

本文选题：GAP-43　切入点：骨髓间充质干细胞　出处：《吉林大学》2009年博士论文

【摘要】： 近年来随着干细胞领域研究的不断深入,为探索神经细胞变性或损伤性疾病的治疗提供了新的思路。骨髓间充质干细胞(BMSCs)具有多向分化潜能,目前已成为成体干细胞领域研究的热点。生长相关蛋白-43(GAP-43)被认为是神经元发育和再生的一个内在决定因子,与神经发育、再生密切相关,因此明确GAP-43在BMSCs向神经样细胞分化过程中的作用对于洞悉干细胞的定向分化机制具有重要意义。本研究分别应用化学诱导剂、生长因子及视网膜条件分化液诱导大鼠BMSCs定向分化为神经样细胞,在此过程中均检测到GAP-43持续性表达,提示其参与了分化过程并可能发挥重要作用。此外,应用基因克隆技术将GAP-43基因克隆并构建到pcDNA3.1(+)载体,转染BMSCs后细胞表达GAP-43,并且向神经样细胞分化能力得到提高,在视网膜条件分化液诱导环境下此种能力提高更加明显。同时,利用RNA干扰技术构建出GAP-43-shRNA表达载体,转染BMSCs后在诱导条件下可特异性抑制GAP-43基因的表达,基因沉默后BMSCs向神经样细胞分化能力受到明显抑制。通过上述研究证明GAP-43为体外培养的BMSCs向神经样细胞分化过程中发挥重要作用的基因之一,表达GAP-43基因将促进BMSCs向神经样细胞分化,抑制该基因表达后细胞分化能力减弱。研究结果对于进一步探索干细胞定向分化机理和可应用于临床的基因治疗方法奠定了理论基础。
[Abstract]:In recent years , with the deepening of research in the field of stem cells , it provides a new idea for exploring the treatment of neural cell degeneration or injury diseases . Bone marrow mesenchymal stem cells ( MSCs ) have the potential to differentiate into adult stem cells . The growth - related protein - 43 ( GAP - 43 ) is considered to be an intrinsic determinant of neuronal development and regeneration , which is closely related to neurodevelopment and regeneration . It is therefore clear that GAP - 43 plays an important role in the differentiation of neural - like cells .

In addition , GAP - 43 gene was cloned and constructed into pcDNA3.1 ( + ) vector . GAP - 43 gene was cloned and transfected into pcDNA3.1 ( + ) vector .

It is proved that GAP - 43 gene plays an important role in the differentiation of neural - like cells in vitro . The expression of GAP - 43 gene will promote the differentiation of bone marrow cells into neural - like cells , and decrease the cell differentiation ability after the gene expression . The results of this study have laid a theoretical foundation for further exploring the mechanism of stem cell differentiation and the application of gene therapy in clinic .

【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2009
【分类号】：R329

【参考文献】