Differentiation Of Mesenchymal Cells Derived From Mouse Fetal Liver Into Islet β-like Cells Cells In Vitro

Objective: To isolate mesenchymal stem cells (MSCs) from fetal liver and induce them differentiate into islet β -like cells. And determine whether these islet β -like cells are able to reverse hyperglycemia in type 1 diabetic mice. Methods: MSCs were isolated from C57BL/6J mouse fetal liver (E11.5-E15.5)by adhesive screening method and induced by high glucose, bFGF(basic Fibroblast Growth Factor), nicotamine in culture medium. The gene expression related to islet β cells such as pancreatic duodenal homeobox-1 (PDX-1) , proinsulin-1 (INS-1) ,glucose transporter-2 (Glut2) was tested by RT-PCR . we stained the clusters with dithizone (DTZ) which is a zinc-chelating agent known to selectively stain pancreatic β cells because of their high zinc content. And Insulin in the treated cells was examined by immunocytochemistry. Finally, we transplanted the treated cells into diabetes mice and monitor the blood glucose in 6 weeks. Results: Under conditions described above, cultures became homogeneous spindle-shaped cells .RT-PCR showed that the treated cells expressed PDX-1, INS-1 and Glut2, while the undifferentiated cells not. After approximately 10 days of treatment, the fetal liver cells formed DTZ-stained cell clusters in flasks (80-120 clusters in a flask 25 cm2 in area). Immunocytochemistry also confirmed that these cell clusters were strongly positive for insulin. The hyperglycemia was reversed in the treated mice.Conclusion: MSCs derived from fetal liver can be induced into islet β -like cells in vitro.These islet β -like cells can synthesize, store and secret insulin. Fetal liver MSCs may be an ideal seed cells for type 1 diabetes.