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目的:从人足月的胎盘羊膜中和Wistar大鼠股骨及胫骨骨髓中分离、培养间充质干细胞(MSCs),研究人胎盘来源MSCs(HPMSCs)和大鼠骨髓来源MSCs(BMSCs)的分离培养方法和生物学特征、表面标志及其多分化潜能。方法:将人足月胎盘组织通过胶原酶Ⅱ消化培养获取HPMSCs,采用全骨髓贴壁法从4周龄Wistar大鼠双侧股骨及胫骨中分离、纯化大鼠BMSCs,运用活细胞计数法检测其增殖能力,采用流式细胞术检测2种细胞表面标志物的表达阳性率;用地塞米松、维生素C和β磷酸甘油诱导其向成骨细胞分化,用茜素红染色鉴定;用胰岛素、地塞米松、IBMX和吲哚美辛诱导其向脂肪细胞分化,以油红O染色鉴定。结果:HPMSCs为梭形贴壁细胞,增殖能力较强,CD44和CD34表达阳性率分别为94.45%和1.67%;BMSCs为圆形、梭形和多角形,增殖能力强,CD44和CD34表达阳性率分别为93.11%和2.68%。2种细胞经过成脂诱导液和成骨诱导液诱导后,茜素红和油红O染色结果均为阳性,表明2种细胞均可向脂肪细胞和成骨细胞分化。结论:HPMSCs与大鼠BMSCs的生物学特征相似,同样具有多分化潜能,HPMSCs具有更强的增殖能力。
OBJECTIVE: To isolate and culture mesenchymal stem cells (MSCs) from human full-term placental amniotic membrane and Wistar rat femur and tibia bone marrow to study the isolation and culture of human placenta-derived MSCs (HPMSCs) and rat bone marrow-derived MSCs (BMSCs) Methods and biological characteristics, surface markers and their potential for differentiation. Methods: Human full-term placenta tissues were obtained by collagenase Ⅱ digestion culture to obtain HPMSCs. Whole bone marrow adherent method was used to separate and purify BMSCs from bilateral femur and tibia of 4-week-old Wistar rats. The viable cell count Proliferation, the positive rate of the expression of the two cell surface markers was detected by flow cytometry; differentiation into osteoblasts was induced by dexamethasone, vitamin C and β-phosphoglycerol, and identified by alizarin red staining; with insulin, Mesotaxel, IBMX and indomethacin induced their differentiation into adipocytes, identified by Oil Red O staining. RESULTS: HPMCCs were spindle-shaped adherent cells with strong proliferative ability, the positive rates of CD44 and CD34 expression were 94.45% and 1.67% respectively. BMSCs were round, spindle and polygonal with strong proliferative ability and positive rate of CD44 and CD34 Respectively 93.11% and 2.68%. After induced by both adipogenic and osteogenic inducing agents, both of the two cells were positive for alizarin red and oil red O staining, indicating that both cells differentiated into adipocytes and osteoblasts. Conclusion: The biological characteristics of HPMSCs and rat BMSCs are similar. They also have the potential of differentiation. HPMSCs have stronger proliferative capacity.