吸脂组织中脂肪干细胞分离和定向分化的研究

Research of isolation and directional differentiation of adipose derived stem cells after suction lipectomy

目的探讨吸脂组织中的干细胞分离和体外诱导分化为表皮样细胞、成骨细胞及脂肪细胞的可能性。方法通过电动负压吸引获取1例行吸脂手术的30岁女性腹部脂肪组织，酶消化法获取脂肪来源干细胞，体外培养扩增，通过流式细胞仪检测表面抗原的表达。取生长良好的第3代人脂肪来源干细胞，分别应用成表皮诱导培养液（70％培养液A＋30％成纤维细胞培养基上清液＋10ng／L表皮生长因子），成骨诱导培养基（DMEM／10％FBS，0．1μmol／L地塞米松，50μmol／L维生素C，10mmol／Lβ-甘油磷酸钠，100U／ml青霉素，100U／ml链霉素）和成脂肪细胞诱导培养基（DMEM＋10％FBS＋500μmol／L1-甲基-3-异丁基黄嘌呤＋1μmol／L吲哚美辛）诱导20d后，分别对成表皮诱导组进行免疫组化检测CK19表达，成骨诱导组进行碱性磷酸酶检测，成脂诱导组进行油红O检测。结果流式细胞仪鉴定结果示，人脂肪来源干细胞CD44和CD49d为阳性，CD34为阴性。诱导20d后，成表皮诱导组示免疫组织化学鉴定结构显示有CK19的表达；成骨诱导组示细胞碱性磷酸酶染色阳性；成脂肪细胞诱导组示油红O染色，胞质内脂滴均被染成红色，证实为脂性液体。结论从吸出的脂肪组织中分离出脂肪来源干细胞，在体外进行了脂肪干细胞的扩增和传代，所分离的脂肪来源干细胞具备多向分化能力。

Objective To observe the formation possibility of epidermic cells, skeletogenous cells and adipose cells by isolation and directional differentiation of adipose derived stem cells （ADSCs） in vitro. Methotis Human abdominal adipose tissues were obtained by vacuum aspiration technique of suction lipectomy sur- gery from a 30-year-old woman. ADSCs were gotten by digestion and amplification. CD antigen was detected by flow cytometry. The ADSCs at 3rd passage were induced in epidermic cells＇ culture solution （70% culture solution A ＋ 30% supernatant fluid from fibroblast medium ＋ 10 ng/LEGF） , skeletogenous cells＇ culture solution （DMEM/10%FBS, 0. 1μmol/L desacort, 50μmol/L vitamin C, 10 mmoL/L sodium β-glycerophosphate, 100 U/ml penicilin, 100 U/mlphytomycin） and adipose cells culture solution （ DMEM ＋ 10% FBS ＋ 500 μmol/L IBMX ＋ 1 μmol/L antinfan） respectively, the expression CK19 in the group which was induced into epidermic cells was detected by IHC, ALP in the group which was induced into skeletogenous cells was detected and the rathonum red in the third group was checked. Results The expression of CD44 and CD49 was positive and CD34 was negative by flow cytometry. The expression of CK19 in the group which was induced into epidermic cells was positive at 20 days after induction, ALP in the group which was induced into skeletogenous cells was positive and the rathonum red in the third group was positive. Condusion Human adipose derived stem cells can be obtained from human adipose tissue after suction lipectomy. The ADSCs can be differentiated into epidermic cells, skeletogenous cells and adipose cells in vitro.