Objective: To establish a method through which murine bone marrow mesenchymal stem cells (MSCs) can be induced into hepatocytes in vitro.
Methods: A conditioned medium of injured hepatocytes (with CCl4 in vivo) was used to culture the isolated MSCs. The differentiated cells were identified by morphological observation, reverse transcription polymerase chain reaction (RT-PCR), immunofluorescence assay (for AFP, Albumin, and CK18) and periodic acid schiff reaction (PAS) for glycogen.
Results: The differentiated cells showed characteristics of hepatocytes. PT-PCR detected AFP mRNA on day 5 and it increased gradually until day 15, and then decreased; CK18 mRNA was detected on day 10; TAT was detected on day 20. Immunofluorescence assay for AFP, albumin and CK18 showed positive staining reactions on day 20. PAS positive glycogen granules appeared in the cytoplasm of the differentiated cells.
Conclusion: MSCs of adult mice cultured in a conditioned medium of injured hepatocytes can differentiate into hepatocytes. This method can be used in further studying of the mechanism of transdifferentiation of MSCs into hepatocytes.