人BMP-2基因转染人脂肪组织源性干细胞诱导体内成骨的实验研究

发布时间：2018-10-14 15:54

【摘要】： 骨缺损是骨科临床常见的难题,现有的治疗方法中有自体骨移植、同种异体骨移植、生物材料充填等,但这些方法都不是理想的治疗模式。而骨组织工程利用具有成骨能力或潜能的种子细胞与生物材料复合,体外培养后植入体内,通过新生的骨组织修复骨缺损,是最理想的治疗模式。以往骨组织工程种子细胞的研究多集中于骨髓间充质干细胞,但其存在数量少、取材困难、体外培养相对困难且易老化、病人不易接受等缺点。而脂肪组织源性干细胞以其自体来源、容易获取、细胞充足、多向分化、基因稳定表达、自体移植无免疫排斥反应、不涉及医学伦理学问题等特点可能成为颇具优势的基因治疗载体。本研究成功从人脂肪组织中分离提取了人脂肪组织源性干细胞。以骨形态发生蛋白基因为目的基因,以人类脂肪组织源性干细胞为靶细胞,进行基因转染,利用转基因技术,结合组织工程种植和培养技术,在体外培养出能稳定生长的、高表达骨形态发生蛋白的转基因人脂肪组织源性干细胞,并研究其体内外成骨能力。证实将组织工程技术与基因治疗技术相结合,可成功将目的基因导入人脂肪组织源性干细胞,经转染后的人脂肪组织源性干细胞在体内表现出良好的成骨能力。本研究对人脂肪组织源性干细胞的成骨潜能和转基因效果做出了客观的、实事求是的评价。为临床实际应用提供了可靠的实验依据。
[Abstract]:Bone defect is a common problem in orthopedic clinic. The existing treatment methods include autologous bone transplantation, allogeneic bone transplantation and biomaterial filling, but these methods are not ideal treatment mode. Bone tissue engineering using seed cells with osteogenic ability or potential to combine with biomaterials is the most ideal treatment mode after cultured in vitro and implanted into the body to repair bone defects through new bone tissue. In the past, the studies of bone tissue engineering seed cells focused on the bone marrow mesenchymal stem cells, but they had some disadvantages, such as small number, difficult to obtain materials, relatively difficult to culture in vitro, easy to aging, and difficult to be accepted by patients. Adipose tissue-derived stem cells are easy to obtain, abundant cells, multidirectional differentiation, stable gene expression, and no immune rejection after autologous transplantation. The characteristics, such as not involving medical ethics, may be an advantageous gene therapy carrier. In this study, human adipose tissue derived stem cells were isolated from human adipose tissue. Taking bone morphogenetic protein gene as target gene and human adipose tissue-derived stem cells as target cells, gene transfection was carried out. Transgenic human adipose tissue derived stem cells with high expression of bone morphogenetic protein and their osteogenic capacity in vitro and in vivo were studied. It was proved that the target gene could be successfully transferred into human adipose tissue derived stem cells by combining tissue engineering with gene therapy. The transfected adipose tissue derived stem cells showed good osteogenic ability in vivo. In this study, the osteogenic potential and transgenic effect of human adipose tissue derived stem cells were evaluated objectively and realistically. It provides reliable experimental basis for clinical application.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2007
【分类号】：R329;R687.3

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