Abstract: Objective To optimize the protocols for isolation and culture of mesenchymal stem cells from rat bone marrow
(BMSCs). Methods BMSCs were isolated by adherence to plastic with frequent medium change and reduced trypsinization
time. The cell growth curves were drawn and the surface markers of BMSCs were detected by flow cytometry. The cells were
induced to differentiate into osteogenic, adipogenic, hepatic and cholic lineages. Results The cells isolated using this method
were positive for CD29, CD44, and CD90 and negative for the hematopoietic surface markers CD45. The osteogenic and
adipogenic differentiation of the BMSCs was verified by alkaline phosphatase staining, Alizarin red staining and Oil red
staining. The cell subcultures up to passage 10 maintained capacities of differentiation into osteogenic and adipogenic lineages.
The BMSCs induced with sequential addition of growth factors, cytokines and hormones differentiated into cells expressing
hepatocyte- and cholangiocyte-specific markers. Conclusion The optimized method allows efficient isolation of homogenous
populations of MSCs from rat bone marrow, which can be induced into multiple cell lineages.