Abstract:Abstract: To investigate the effects of unsaturated fatty acids on the morphology, proliferation, immunophe-notype and differentiation potential of human adipose mesenchymal stem cells (ADSCs), ADSCs were isolated from subcutaneous adipose tissue by digestion with 0.15% type Ⅷ collagenase. After subculture, the cells were cultured with oleic acid and linolenic acid, respectively, and were observed by inverted microscope. The cell growth curve was generated by cell-counting. The flow cytometry was used to examine the expres-sion of the surface markers. After being cultured with the differentiation medium, the cells were stained by oil red O for adipogenesis, alizarin red for osteogenesis and alcian blue for chondrogenesis. Meanwhile, quantitative PCR analysis was used to determine the relative mRNA expression levels of stem cell-related genes 9 days after induction. The results showed that ADSCs with uniform morphology could be obtained by using 0.15% type Ⅷ collagenase. Compared with the control group, the groups cultured with 20 μmol/L oleic acid or linolenic acid had about 20% increase in the number of ADSCs within the same amount of time. The flow cytometry examination showed that expressions of CD13, CD29, CD44, CD73, CD90 and CD105 were positive, and those of CD31 and CD45 were negative. The cells successfully differentiated into adipogenic, osteogenic and chondrogenic lineages after cultured with 20 μmol/L oleic acid or linolenic acid, and were stained positively with oil red O, alizarin red and alcian blue. The expression levels of adipogenic gene PPAR-γ, chondrogenic genes SOX9, COL2A1, and osteogenic genes ALP, OCN, RUNX2 were significantly higher compared with those in the control group. The results showed that oleic acid and linolenic acid could promote ADSC proliferation, maintain their immunophenotype and improve their differentiation ability.
引用本文:
李锦灵, 林立龙, 李治寰. 不饱和脂肪酸对人脂肪间充质干细胞生物学特性的影响[J]. 生命科学研究, 2021, 25(1): 15-23.
LI Jin-ling, LIN Li-long, LI Zhi-huan. Effects of Unsaturated Fatty Acids on Biological Characteristics of Human Adipose Mesenchymal Stem Cells. Life Science Research, 2021, 25(1): 15-23.