脂肪来源具有干细胞潜能的细胞的体外生长特性

发布时间：2018-01-17 23:24

本文关键词：脂肪来源具有干细胞潜能的细胞的体外生长特性　出处：《浙江大学》2006年硕士论文　论文类型：学位论文

【摘要】：【目的】:对脂肪来源的具有干细胞潜能的细胞进行体外培养,并对此类细胞体外生长的特性进行研究,为进一步的研究做基础。【材料与方法】:利用整形外科成熟的负压吸脂术获取脂肪组织,经过一定处理后以酶解法得到具有增殖分化能力的幼稚细胞,此类细胞被称为PLA(Processed Lipoaspirate)细胞,而后在体外培养扩增。利用β-半乳糖苷酶染色法测定PLA的衰老率,利用流式细胞仪分析培养细胞的表面标记物;一部分细胞放置于半固体培养基中培养,利用油红染色检测细胞类型。【结果】:在脂肪组织中确实存在能不断增殖的未成熟细胞,多次传代后仍保留增殖能力,第3代细胞的衰老率为:0.357±0.0178,而第15代细胞的衰老率为:0.371+0.0240(p0.05,无明显差异),形态学及表面标记物均与骨髓间充质干细胞(Mesenchymal stem cell, MSC)类似,通过流式细胞技术(Fluorescence-activated cell sorter, FACS)鉴定,PLA细胞表面黏附分子CD44、CD29均高表达,而造血干细胞调节因子CD34、CD45均低表达。半固体凝胶培养未能促进组织分化。【结论】:脂肪组织中确实存在一群具有极强增殖潜能的细胞,在多次分裂后,仍保持其增殖潜能。同时此类原始细胞的细胞表型与骨髓
[Abstract]:Objective: to culture adipose cells with stem cell potential in vitro and to study the characteristics of their growth in vitro. [materials and methods]: mature vacuum liposuction was used to obtain adipose tissue. After certain treatment, immature cells with proliferative and differentiation ability were obtained by enzymatic hydrolysis. These cells were called PLA(Processed lipoaspirate cells and were cultured and amplified in vitro. The senescence rate of PLA was determined by 尾 -galactosidase staining. The surface markers of cultured cells were analyzed by flow cytometry. Some of the cells were cultured in semisolid medium and the cell types were detected by oil red staining. [results]: there were immature cells in adipose tissue which could proliferate continuously, and the ability of proliferation remained after multiple passages. The senescence rate of the third passage cells was: 0.357 卤0.0178. However, the senescence rate of the 15th passage cells was 0. 371 0. 0240 (p 0. 05). Morphological and surface markers were similar to mesenchymal stem cells (MSCs). The surface adhesion molecule (CD44) of PLA cells was identified by flow cytometry (FACSs) and fluorescence-activated cell sorter (FACSs). CD29 was highly expressed, while CD34 / CD45, a hematopoietic stem cell regulator, was low. Semisolid gel culture did not promote tissue differentiation. [conclusion]: there is indeed a group of cells with strong proliferative potential in adipose tissue, which remain proliferative potential after multiple division. Meanwhile, the phenotypes and bone marrow of these primitive cells are also present.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R329.2

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