大鼠脂肪干细胞体外诱导成平滑肌样细胞的实验研究

发布时间：2018-01-05 11:32

本文关键词：大鼠脂肪干细胞体外诱导成平滑肌样细胞的实验研究　出处：《华中科技大学》2010年硕士论文　论文类型：学位论文

【摘要】：【目的】研究大鼠脂肪干细胞(ADSCs)体外单层培养条件下诱导分化为平滑肌样细胞的可行性。【方法】从SD大鼠的腹股沟脂肪垫分离获取脂肪干细胞,测定其生长曲线。取第4代细胞用成脂诱导液诱导分化,并用油红O染色鉴定。取第4代细胞用成骨诱导液诱导分化,并用Von Kossa染色鉴定。取第4代细胞用含有β-巯基乙醇的成平滑肌诱导液诱导,并用免疫组化的方法检测α平滑肌肌动蛋白(α-SMA)的表达。【结果】脂肪干细胞成梭形和多角形,体外生长迅速,生长曲线表明传代2天后细胞进入对数生长期。第4代脂肪干细胞,成脂诱导后,油红O染色证实细胞内存在脂滴;成骨诱导后,Von Kossa染色证实有钙化结节。脂肪干细胞诱导平滑肌样细胞免疫组化结果,含β-巯基乙醇诱导液诱导组和未诱导组细胞α-SMA的表达阳性率分别为(29.80±6.89)%、(2.89±1.24)%。诱导组细胞α-SMA的表达阳性率高于未诱导组,差异有统计学意义(P0.01)。【结论】脂肪干细胞经诱导后出现明显的平滑肌细胞特征,可成为平滑肌相关疾病在组织工程研究上新的种子细胞来源。
[Abstract]:[purpose] To study the feasibility of differentiation into smooth muscle like cells (SMC) induced by rat adipose stem cells (ADSCs) in vitro monolayer culture. [methods] The adipose stem cells were isolated from the groin fat pad of SD rats and their growth curves were measured. The differentiation of the cells was induced by osteoblast inducer and Von Kossa staining. The fourth passage cells were induced by 尾 -mercaptoethanol. The expression of 伪-smooth muscle actin (伪 -SMA) was detected by immunohistochemistry. [results] Adipose stem cells were spindle-shaped and polygonal, and grew rapidly in vitro. The growth curve showed that the cells entered logarithmic growth phase 2 days after passage, and the fourth generation of adipose stem cells was induced by adipogenesis. Oil red O staining confirmed the presence of lipid droplets in the cells. Calcified nodules were confirmed by Von Kossa staining after osteogenic induction. Immunohistochemical results of smooth muscle like cells induced by adipose stem cells were observed. The positive rate of 伪 -SMA expression in cells induced by 尾 -mercaptoethanol was 29.80 卤6.89%, respectively. The positive rate of 伪 -SMA expression in the induction group was higher than that in the uninduced group, and the difference was statistically significant (P 0.01). [conclusion] After induction, adipose stem cells have obvious characteristics of smooth muscle cells, which may be a new source of seed cells for tissue engineering of smooth muscle related diseases.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R329

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