Objective To observe morphology.identify cell surface CD marker and test the ability of proliferation and differentiation of adipose derived stem/stromal cells (ASCs) which were isolated from intact fat ex vivo. Methods ASCs were isolated by collagenase digesting from intact fat of abdomen harvesting of skin grafts. The 3rd passage ASCs were applied to do the experiments: HE staining, flow cytometry.MTT method and cell cycle testing. Besides.passage 3 cells were induced to differentiate into lipocytes and osteocytes. Oil red staining was applied to examine adipogenic induction. Alizarin red staining was applied to examine osteogenic induction. Results Primary ASCs were multi-angular and short spindle cells at the first culture fluid changing. HE staining showed that Passage 3 cells grew like swirls and were long spindle-shaped. Row cytometry showed CD29+,CD31-,CD34-,CD44t,CD45-,CD49+,CD106-,CD133-.MTT manifested the growth and proliferation of ASCs was great. Cell cycle testing showed ASCs in G1 period accounted for 86.8% and G2 period 8.77%. Oil red staining and alizarin red staining both demonstrated positive reactions but not in control groups. Conclusion Human ASCs were a kind of mesenchymal stem cells with great application prospect, which characterized adherent growth.multipotent differentiation and stem cell phenotype.%目的：在体外从脂肪块中分离出脂肪干细胞(adipose derived stem/stromal cells，ASCs)，对其进行形态观察、干细胞鉴定、增殖和分化能力检测。方法：将腹部取皮术的皮下脂肪利用胶原酶消化法，进行体外分离培养，取第3代的细胞进行细胞爬片HE染色、流式鉴定、MTT、细胞周期检测等，利用成脂和成骨培养液诱导，油红0和茜素红染色鉴定诱导结果。结果：原代培养第1次换液时细胞多呈多角形和短梭形，第3代ASCs细胞爬片HE染色显示形态多为长梭形，呈漩涡状生长；流式鉴定显示：CD29+，CD31-，CD34-，CD44+，CD45-，CD49+，CD106-，CD1 33-;MTT显示ASCs生长增殖活性强；细胞周期检测结果显示：G1=86.8％，G2=8.77％；成脂诱导后油红0染色阳性，对照组为阴性；成骨诱导后茜素红染色阳性，对照组为阴性。结论：人ASCs具有贴壁生长、多向分化以及干细胞表型等特征，且生长增殖活性强，是一种很有应用前景的间充质干细胞。
展开▼
