大鼠脂肪干细胞分离培养及成脂成骨分化潜能的研究
发布时间：2018-09-02 09:02
【摘要】： 目的:研究脂肪干细胞(adipose tissue-derived stem cells,ADSCs)在体外分离培养和在一定条件下诱导分化为脂肪细胞和成骨细胞的能力,探讨其作为骨组织工程细胞的可行性。 方法:无菌切取SD大鼠腹股沟处的皮下脂肪,Ⅰ型胶原酶消化法获取原代脂肪干细胞,接种至含10%胎牛血清的DMEM培养液,培养并适时传代,每日在倒置显微镜下观察并记录细胞形态和增殖特征。第3代细胞分别在成脂成骨诱导液中诱导分化,并采用油红染色,碱性磷酸酶(ALP)染色,Von Kossa染色进行鉴定。 结果:从大鼠脂肪组织中分离出的脂肪干细胞,在体外呈成纤维细胞样迅速增殖,可在异丁基-甲基黄嘌呤、地塞米松、胰岛素等诱导下,表现出脂肪细胞特性;可以在地塞米松、抗坏血酸、β—甘油磷酸钠的诱导下,表现出成骨细胞特性,该细胞碱性磷酸酶(ALP)染色ALP活性显著增高,Von Kossa染色出现矿化结节。 结论:ADSCs易于在体外分离培养,并扩增迅速,其生物学特性稳定,经成脂诱导可分化为脂肪细胞,经成骨诱导可分化为成骨细胞,可作为骨组织工程的种子细胞。
[Abstract]:Aim: to study the ability of adipose stem cells (adipose tissue-derived stem cells,ADSCs) to differentiate into adipocytes and osteoblasts in vitro and to explore its feasibility as bone tissue engineering cells. Methods: the subcutaneous fat of the groin of SD rats was cut aseptically and the primary adipose stem cells were obtained by type I collagenase digestion. The primary adipose stem cells were inoculated into the DMEM culture medium containing 10% fetal bovine serum. Cell morphology and proliferation were observed and recorded daily under inverted microscope. The third generation cells were induced to differentiate in adipogenic osteoblast and identified by oil red staining and alkaline phosphatase (ALP) staining with von Kossa staining. Results: adipose stem cells isolated from adipose tissue of rats proliferated rapidly as fibroblasts in vitro, and showed adipocyte characteristics induced by isobutyl-methylxanthine, dexamethasone, insulin and so on. The osteoblasts could be induced by dexamethasone, ascorbic acid and 尾 -glycerophosphate. The activity of ALP by alkaline phosphatase (ALP) staining was significantly higher than that by Von Kossa staining. Conclusion it is easy to isolate and culture in vitro and expand rapidly, and its biological characteristics are stable. The adipocytes can be differentiated into adipocytes by adipogenic induction and osteoblasts can be differentiated into osteoblasts by osteogenesis, which can be used as seed cells for bone tissue engineering.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R329
本文编号：2218884
[Abstract]:Aim: to study the ability of adipose stem cells (adipose tissue-derived stem cells,ADSCs) to differentiate into adipocytes and osteoblasts in vitro and to explore its feasibility as bone tissue engineering cells. Methods: the subcutaneous fat of the groin of SD rats was cut aseptically and the primary adipose stem cells were obtained by type I collagenase digestion. The primary adipose stem cells were inoculated into the DMEM culture medium containing 10% fetal bovine serum. Cell morphology and proliferation were observed and recorded daily under inverted microscope. The third generation cells were induced to differentiate in adipogenic osteoblast and identified by oil red staining and alkaline phosphatase (ALP) staining with von Kossa staining. Results: adipose stem cells isolated from adipose tissue of rats proliferated rapidly as fibroblasts in vitro, and showed adipocyte characteristics induced by isobutyl-methylxanthine, dexamethasone, insulin and so on. The osteoblasts could be induced by dexamethasone, ascorbic acid and 尾 -glycerophosphate. The activity of ALP by alkaline phosphatase (ALP) staining was significantly higher than that by Von Kossa staining. Conclusion it is easy to isolate and culture in vitro and expand rapidly, and its biological characteristics are stable. The adipocytes can be differentiated into adipocytes by adipogenic induction and osteoblasts can be differentiated into osteoblasts by osteogenesis, which can be used as seed cells for bone tissue engineering.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R329
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相关期刊论文 前3条
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