Optimal induction conditions for adipose-derived stem cells differentiating into lymphatic endothelial cells / 中国组织工程研究

ABSTRACT

BACKGROUND:Our previous studies have shown that adipose-derived stem cels under vascular endothelial growth factor C (VEGF-C) can be induced to differentiate into lymphatic endothelial cels that are confirmed by lymphatic vascular endothelial hyaluronan receptor-1 staining. However, its optimal induction program is not clear. OBJECTIVE:To investigate the best condition for the differentiation of adipose-derived stem cels into lymphatic endothelial cels under induction of VEGF-C156s. METHODS: Adipose tissues from healthy adults were colected to isolate adipose-derived mesenchymal stem cels using trypsin digestion method. Flow cytometry was employed to detect cel surface markers, andin vitro differentiation capacity was identified by adipogenic and osteogenic induction. Passage 3 cels at good growth state were selected and divided into six groups cels in control group were cultured in low-glucose DMEM, and those in the rest five groups were treated with 25, 50, 100, 200, 300 μg/L VEGF-C156s, respectively. RESULTS AND CONCLUSION:Adipose-derived stem cels were successfuly obtained by trypsin digestion and purification, and then differentiated into lymphatic endothelial cels under the induction of VEGF-C156s, basic fibroblast growth factor and other growth factors. No cels were positive for lymphatic vascular endothelial hyaluronan receptor-1 in the control group. After 8 days of induction, few cels were positive in the 25 μg/L VEGF-C156s group; a great amount of positive cels were visible in the 50 and 100 μg/L VEGF-C156s groups; 200 and 300 μg/L VEGF-C156s resulted in a large number of deaths in the cels. These findings indicate that it is optimal for adipose-derived stem cels to differentiate into lymphatic endothelial cels under 8-day induction of 50 μg/L VEGF-C156s.