The Effects Of Ca²⁺ On The Differentiation Of Bone Marrow Stromal Stem Cells Into Hepatocytes

Objectives The differentiation of Bone marrow stromal stem cells (BMSCs) into hepatocytes has a great potential use in regenerative medicine and tissue engineering like treating liver damages. The differentiation of mice BMSCs into hepatocytes had been studied, however the mechanism of BMSCs into hepatocytes is unclear. In this study the role of Ca2+in the differentiation of BMSCs into hepatocytes in vitro was detected.Materials and Methods1. MaterialsHuman lung adenocarcinoma A549cells (Medical Center of Affiliated Hospital of Qingdao University, Medical College) DMEM and fetal bovine serum (Hyclone) Alpha antitrypsin (AAT) and albumin polyclone antibody (Abcam) Berta nerve growth factor (β-NGF) and Indocyanine green (ICG)(Sigma) Fluo-3/AM (Biotium Inc) Nimodipine (Yunnan supertrack bio pharmaceutical)2. MethodsBMSCs were isolated from bone marrow of mice and cultured in low-glucose DMEM with20%FCS for expanding. Then BMSCs at3rd generation were respectively applied with the hepatocyte abstraction (G,500μg/ml) and β-NGF from rat (50ng/ml) for20days and β-NGF (50ng/ml) by combining Nimodipine. The features of differentiatied hepatocytes were identified by immunocytochemistry for the exclusive marker of hepatocytes alpha antitrypsin (AAT) and albumin (ALB) respectively, and by ICG for the hepatocytes transferring function. The proliferation of BMSCs treated with Nimodipine was detected by MTT.3. Results(1) The differentiated cells from BMSCs at3rd generation induced with hepatocyte abstraction and β-NGF from rat respectively were showed round or oval in shape, ICG positive and positive expression of AAT and ALB proteins compared with control groups(P<0.05).(2)[Ca2+] i of the hepatocytes from BMSCs was higher than control group(P<0.05).(3) Nimodipine could block BMSCs proliferation and differentiation into hepatocytes.4. Conclusions (1) The cytokine G and P-NGF HGF could induce BMSCs differentiation into functional hepatocytes.(2) Ca2+played a significant role in the proliferation and differentiation of BMSCs.