Abstract:

BACKGROUND: Pancreatic ductal epithelium cells (PDECs) can differentiate into insulin-producing cells in vitro or in vivo, but the cells have poor proliferation in vitro. Thus, the cell transplantation therapy for diabetes is limited.
OBJECTIVE: To investigate the effect of keratinocyte growth factor (KGF) at different concentrations on the proliferation of PDECs in order to seek the optimal serum dose.
METHODS: The PDECs of rats were isolated by injecting collagenase Ⅴ solution and purified by removing the islets with Ficoll density gradient centrifugation. Then the PDECs were identified by immunocytochemical staining and RT-PCR. Different concentrations of KGF were added into medium. The proliferation ability of PDECs was determined by cells counting and MTT assay.
RESULTS AND CONCLUSION: Immunocytochemical staining results showed positive expression of CK19 in PDECs; Morphology of the PDECs in the different groups were similar. The PDECs numbers of the 5 μg/L KGF group were significantly lower than that of the other KGF groups (P < 0.01), and the proliferation of the groups cultured with 10, 15 and 20 μg/L KGF were similar (P > 0.05). The KGF at different concentrations can significantly stimulate the proliferation of PDECs, the optimized concentration is 10 μg/L．