Growth factor-free cultured rat bone marrow derived mesenchymal stem cells towards hepatic progenitor cell differentiation

Abstract

Induction of mesenchymal stem cells (MSCs) differentiation by growth factors is not likely to be acceptable for clinical application. In this study, MSCs and hepatocytes isolated from male Sprague-Dawley rats were indirectly co-cultured by sharing media, without any growth factor, for 14 days. Differentiation was investigated by detection of hepatic markers, albumin, α-fetoprotein (AFP), cytokeratin-18 (CK18), and bile ductal epithelial cell marker cytokeratin-19 (CK19) on reverse transcription polymerase chain reaction (RT-PCR), immunofluorescence staining, and Western blot. Periodic acid-schiff (PAS) staining, low density lipoproteins (LDL) uptake analysis, and urea assay were performed to detect metabolic activities of differentiated cells. MSCs in co-culture group showed positive expression for albumin, AFP, and CK19, but little for CK18. Data indicates that co-cultured MSCs were differentiated into oval cell stage, the hepatic progenitor, but not directly toward complete hepatocytes. Our finding suggests that indirect co-culture system has potential to generate regenerative hepatocytes by hepatic progenitor cells differentiated from MSCs