摘要
目的探索人脂肪组织来源干细胞体外培养的最佳条件。方法人腹部皮下脂肪抽吸取材,采用胶原酶消化法获取细胞,采用含血清的DMEM培养液体外培养细胞。比较了不同胶原酶浓度、胶原酶消化时间和血清浓度对于人脂肪来源干细胞体外培养的不同效果。结果采用0.5、1.0、1.5、2.0mg/ml四种不同胶原酶浓度进行消化,发现在消化时间为1h时,2.0mg/ml组消化细胞总数最多,活细胞比值与其他组比较,差异有显著意义。在消化时间为2h时,消化细胞总数较1h各组均增加,同时,1.0、1.5和2.0mg/ml组消化细胞总数无统计学差异,活细胞比值1.0mg/ml组和1.5mg/ml组最高。采用0、5%、10%、15%、20%五种不同血清浓度培养后发现,15%和20%组生长特性最佳,明显优于另外三组。结论对于人脂肪组织来源干细胞,胶原酶浓度1.0mg/ml、消化时间2h、血清浓度15%是最佳的培养条件。
Abstract： Objective To explore the optimum condition for human adipose- derived stem cells（hADSC） culture in vitro. Methods Adipose tissues were harvested by liposuction from the abdominal wall and the collagenase digestion method was used for cell separation. Dulbecco＇s Modified Eagle Medium（DMEM） with fetal bovine serum（FBS） was used for cell culture. Collagenase at different concentrations and durations and FCS with different concentrations were used. Results ①Based on the collagenase concentration, cells were divided into four groups, with concentrations of 0.5,1.0,1.5,2.0 mg/ml. At a duration of 1 hour, the 2.0 mg/ml group had the highest total cell number（TCN） and the living cell rate（LCR） was statistically different from the other groups. When the duration was 2 hours, TCN increased in every group and there were no statistical differences among the 1.0,1.5,2.0 mg/ml groups. LCR was the highest in the 1.0 mg/ml and 1.5 mg/ml groups. ②Based on FBS concentration, the cells were divided into five groups, including 0, 5%, 10%, 15% and 20% groups. The growth characteristics were the best in the 15% and 20 % groups. Conclusion The optimal culture conditions for human adipose - derived stem cells are collagenase concentration at 1.0 mg/ml, with digestion duration of 2 hour and FBS concentration of 15%.
出处
《中国美容整形外科杂志》
CAS
2007年第3期223-226,共4页
Chinese Journal of Aesthetic and Plastic Surgery
关键词
脂肪来源干细胞
体外培养
优化
Adipose - derived stem cell
Culture in vitro
Optimization
