The evaluation of function and ultramicrostructure for the hepatocyte-like cells derivated from fetal marrow mesenchymal stem cells / 中华传染病杂志

ABSTRACT

Objective To evaluate the hepatocyte function and ultramicrostructure of marrow mesenchymal stem cells (MMSCs) derivation hepatocyte like cells. Methods MMSCs were isolated from fetal marrow. MMSCs were cultured and induced in vitro in 1% Matrigel as matrix, 2.5 ?mol/ml azacitidine (AZA) pretreatment for 10～12 h, hepatocyte growth factor (HGF) 10 ng/ml+fibroblast growth factor 4 (FGF4) 10 ng/ml+hepatocyte growth medium (HGM). Albumin (ALB) level in culture supernatant was determined with enzyme linked immunosorbent assay (ELISA), capability for dealing with ammonia into urea was detected with urease method, staining for glycogen of undifferentiated MMSCs and MMSCs derivated hepatocyte like cell at induction days 7, 14, 21 and 28 was conducted with periodic acid Schiff's (PAS) test, ultramicrostructure of MMSCs was observed by electromicroscope. Results Undifferentiated MMSCs did not produce ALB and urea. Following treatment with FGF4 and HGF, ALB and urea production by induced MMSCs increased in a time dependent manner. Glycogen storage was first seen by day 14, and maximum levels were seen after day 28, enhancement of glycogen staining for induced MMSCs showed that hepatocyte like cell had glycogen synthesis capability. Undifferentiated MMSCs possessed the construction features of juvenile cell. Induced MMSCs at day 28 had differentiated and had the polarity construction features of epithelioid cell. Conclusions The induced MMSCs has hepatocyte specific construction and functional features.