摘要
目的:对同一个体不同部位来源的脂肪干细胞体外成脂诱导分化的能力进行比较,为脂肪干细胞的进一步应用提供实验依据。方法:实验于2004-11/2005-05在中国协和医科大学整形外科医院研究中心完成。提取5例女性临床吸脂患者(平均年龄32岁)8个部位(大腿前侧、臀上部;臀上部、上腹部;髂腰部;上腹部;背部、上臂)脂肪抽吸组织中的间充质干细胞,体外进行成脂诱导分化。在对照培养基(含体积分数0.1胎牛血清的DMEM培养基)中培养的第1代脂肪干细胞,以3×104细胞/皿接种到35mm培养皿中,将对照培养基更换为成脂诱导培养基,进行成脂诱导,每周换液2次,以加入对照培养基的培养皿设为阴性对照。继续培养2周,分别取诱导后的细胞和阴性对照细胞各两皿进行油红O染色,以证实细胞中有无脂滴形成。通过油红O染色及阳性细胞记数,分别对来源于3例患者的2个不同部位的脂肪抽吸标本中的脂肪干细胞向成脂细胞分化的能力进行定性和半定量检测,采用卡方检验对不同细胞的成脂诱导分化率进行测定。结果:①脂肪干细胞成脂诱导分化能力的测定结果:从不同个体的每100mL脂肪抽吸组织中提取的平均脂肪干细胞数目有所不同,从2.45×108个/100mL～1.25×109个/100mL不等;但从同一个体不同部位的脂肪中所获得的干细胞数目却比较近似;根据各例细胞成脂诱导分化率,可得出5例患者8个部位细胞的平均成脂诱导分化率为36.2%。经统计学分析证实同一个体不同部位来源的细胞具有不同的成脂分化能力,获得的平均干细胞数目与年龄无关(r=0.098,P=0.817),而细胞的成脂分化能力与年龄呈明显负相关(r=-0.568,P<0.01)。②成脂诱导后脂肪干细胞形态变化及油红O染色定性结果。脂肪干细胞经诱导后体积增大,分化为成脂细胞,细胞浆中可见许多大小不等的橙红色脂滴,多者可占整个细胞体积的80%～90%
AIM： To compare the adipogenic differentiation ability in vitro of induced human adipose stem cells from different sites of the same body, so as to provide experimental foundation for further application of adipose stem cells （ASCs）.
METHODS： The experiment was carried out in the Research Center of Plastic Surgery Hospital, Peking Union Medical College from November 2004 to May 2005. The mesonchymal stem cells in lipe^pirates were extracted from 8 sites （anterior thigh, upper buttocks; upper buttocks, upper abdomen; iliolumbalis; upper abdomen; back, upper arm） of 5 female donors （mean age： 32 years old） undergoing the lipesuction in clinic, then these cells were induced to differentiate to the adipegenic cells in vitro. ASCs at passage 1, cultured in the control media （DMEM, supplemented with 10% FBS） were plated at a density of 3×lO^-4 cells every dish into 35 mm culture dishes, and the control media was replaced with adipogenic media （AM）. The media was changed twice weekly and the culture dish added with the control med.ia served as negative control. Two weeks later, the.induced cells and negative cells were assessed with an Oil. Red-O stain to identify whether there was lipid in cells or not. Then through the Oil Red O stain and pesitive cell counting, the adipogenie differentiation ability of induced human adipose stem cells from 2 different sites of 3 patients were assessed with qualitative and send-quantitative analysis. The adipegenie differentiation rates of different cells were determined with ehisquare test.
RESULTS： ①The measured results of adipogenie differentiation ability of ASCs： The mean number of eells isolated from per 100 mL of lipeaspirates of different donors were quite different, the mean cell yields were ran .ged from 2.45×10^8/100 mL to 1.25×10^-9/100 mL; but the mean number from different sites of the same body were quite similar; the mean adipegenic differentiation rate measured from 8 sites of 5 donors was 36.2% calculated according to the adipeg
出处
《中国临床康复》
CSCD
北大核心
2006年第29期44-46,i0002,共4页
Chinese Journal of Clinical Rehabilitation
