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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 d后细胞进入对数生长期。第4代细胞成脂诱导后,油红O染色证实细胞内存在脂滴;成骨诱导后,Von Kossa染色证实有矿化结节。脂肪干细胞诱导平滑肌样细胞免疫组化结果,β-巯基乙醇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(P<0.01)。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。
AIM: To investigate the feasibility of inducing the differentiation of rat adipose derived stem cells (ADSCs) into smooth muscle-like cells under monolayer culture in vitro. Methods: Adipose-derived stem cells were isolated from the inguinal fat pad of SD rats and their growth curves were measured. Generation 4 cells were induced to differentiate with adipogenic inducer and identified by Oil Red O staining. Generation 4 cells were induced to differentiate using osteogenic induction fluid and identified by Von Kossa staining. The 4th generation cells were induced with smooth muscle inducing fluid containing β-mercaptoethanol and the expression of α-smooth muscle actin (α-SMA) was detected by immunohistochemistry. Results: Adipose-derived stem cells became fusiform and polygonal, and grew rapidly in vitro. The growth curves showed that the cells entered the logarithmic growth phase after 2 days of passage. After passage 4 adipogenic induction, oil red O staining confirmed the presence of lipid droplets in the cells; Von Kossa staining confirmed mineralized nodules following osteogenic induction. The positive rate of α-SMA in β-mercaptoethanol-induced and non-induced cells was (29.80 ± 6.89)% and (2.89 ± 1.24)%, respectively. The positive rate of α-SMA in induction group was higher than that in non-induction group (P <0.01). CONCLUSION: Adipose-derived stem cells show obvious characteristics of smooth muscle cells after induction, which may be a new source of seed cells in tissue engineering of smooth muscle-related diseases.