扩增后脂肪源干细胞的成骨分化及体外免疫调节的研究

发布时间：2018-11-13 06:55

【摘要】： 目的:探讨人脂肪源干细胞( adipose derived stem cells,ADSCs)在体外分离、培养、扩增的方法及扩增后向成骨细胞诱导分化与免疫调节的能力。方法:以组织消化的方法分离脂肪组织中的干细胞体外扩增培养,扩增后的ADSCs在成骨诱导条件下促进其向成骨细胞的分化,以明确其分化能力。同时检测扩增后的ADSCs在体外抑制淋巴细胞增殖的能力,以评价ADSCs的免疫原性及免疫调节功能。结果:体外分离培养的脂肪源干细胞生长稳定,扩增速度快。流式细胞仪检测后显示高表达干细胞相关抗原。向成骨细胞诱导后经免疫组化染色及RT-PCR分析显示诱导的细胞分泌成骨细胞的特异性细胞外基质并形成钙结节。同时扩增后的ADSCs在体外不会引起淋巴细胞的增殖并且在混合淋巴细胞反应中可以抑制淋巴细胞的增殖。结论:脂肪源干细胞在体外分离培养简单,扩增速度快,并且扩增后仍具有定向成骨分化及免疫调节的能力,是可靠的组织修复和细胞治疗的种子细胞来源。
[Abstract]:Aim: to investigate the methods of isolation, culture and amplification of human adipose stem cells (adipose derived stem cells,ADSCs) in vitro and their ability to induce differentiation and immunomodulation into osteoblasts. Methods: stem cells from adipose tissue were isolated by tissue digestion and cultured in vitro. The expanded ADSCs promoted the differentiation of adipose cells into osteoblasts under the condition of osteogenesis. In order to evaluate the immunogenicity and immunomodulatory function of ADSCs, the ability of expanded ADSCs to inhibit lymphocyte proliferation in vitro was also detected. Results: the adipose stem cells isolated and cultured in vitro grew stably and expanded quickly. High expression of stem cell associated antigen was detected by flow cytometry. After induction to osteoblasts, immunohistochemical staining and RT-PCR analysis showed that the induced cells secreted the specific extracellular matrix of osteoblasts and formed calcium nodules. Simultaneously amplified ADSCs did not induce lymphocyte proliferation in vitro and inhibited lymphocyte proliferation in mixed lymphocyte reaction. Conclusion: adipose-derived stem cells are easy to be isolated and cultured in vitro and have the ability of directional osteogenesis differentiation and immunomodulation, which is a reliable source of seed cells for tissue repair and cell therapy.
【学位授予单位】：新疆医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R329

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