HGF and G-CSF induce the differentiation of rat bone marrow mesenchymal stem cells into hepatocyte-like cells

Objective To investigate if the hepatocyte growth factor(HGF) and granlocyte-colony stimulating factor(GCSF) induce differentiation of rat bone marrow mesenchymal stem cells(BM-MSCs) into hepatocyte-like cells.Methods The BM-MSCs were isolated and cultured by adherent method,then identif;% 96% 94ied by flow cytometry.The BMMSCs of the third passage were used as four groups-control group(10% FBS) and intervation groups(0.1 μmol/L G-CSF;20 ng/mL HGF;0.1 μmol/L G-CSF + 20 ng/mL HGF).The mRNA and protein were extracted on 7,14,21 days,then detected by RT-PCR and Western blot.Results The third passage BM-MSCs of intervation groups(20 ng/mL HGF and 0.1 μmol/L G-CSF +20 ng/mL HGF) got hepatocyte like feafure after induction.The results tested by flow cytometry analysis were as follows: CD34-PE 0.3%,CD45-FITC 0.1 %,CD90-FITC 99.6 %,CD105-PE 99.8 %.The mRNA and protein of ALB and AFP were detected respetively by RT-PCR and Western blot on day 7,14,and 21 from inervation groups(10% FBS + 20 ng/mL HGF;10% FBS + 0.1 μmol/L G-CSF + 20 ng/mL HGF),however,the amount of ALB increased as time went by,while AFP decreased to extremely low on day 21.Meaningfully,the difference between the two groups is significant(P 0.05).Conclusions HGF can induce the differentiation of BM-MSCs into hepatocyte-like cells,which can be significantly enhanced by G-CSF.