Abstract
Background:We evaluated(1) the potential differentiation ability of bone mesenchymal stromal cells(BMSCs) into pancreatic lineage cells on rat acellular pancreatic bioscaffold (APB) and (2) the effect of differentiated BMSCs on chronic pancreatitis in vivo.Methods:After BMSCs were isolated and characterized, they were dynamically cultured on APB and statically cultured in tissue culture flasks (TCFs), with or without growth factor(GF) in both culture systems. We assessed cytological behavior, such as the proliferation and differentiation of BMSCs, by morphological observation, flow cytometry, enzyme-linked immunosorbent assay (ELISA), quantitative real-time/reverse transcriptase polymerase chain reaction, and Western blot analysis. For the in vivo study, we evaluated the pancreatic fibrosis and pathological scores.We detected the expression of α-SMA,collagen types I and III,and IL-10 in pancreatic tissue by ELISA.Results:The most appropriate flow rate for the dynamic culture of BMSCs was 4mL/min. The proliferation rates of BMSCs in the APB groups were significantly higher than in the TCF groups.During the pancreatic lineage cell differentiation process, APB induced BMSCs to express mRNA markers such as PDX-1 and PTF-1 at higher levels. In contrast, the marker Oct4 was expressed at a lower level in the APB group. All tested pancreatic cytokeratins, including α-Amy, CK7, Flk-1, and C-peptide, were expressed at higher levels in the APB group. The secretion of metabolic enzymes, such as Amy and insulin, was higher in the APB system. By scanning electron microscopy and transmission electron microscopy, the ultrastructure of BMSCs in the APB group further revealed the morphological characteristics of pancreatic-like cells. In the in vivo study, the expression of α-SMA and collagen types I and III in tissues was significantly lower in differentiated BMSCs group, whereas the levels of IL-10 in pancreatic tissue were higher in differentiated BMSCs with significant difference. In addition, in both the in vitro and the in vivo study, GF significantly improved proliferation, differentiation, and pancreatic cell therapy.Conclusion: Our data showed (1) the capacity of APB, a three-dimensional pancreatic biomatrix, to promote BMSC differentiation toward pancreatic lineage and pancreatic-like phenotypes, and (2) the considerable potential of using these cells for pancreatic cell therapies and tissue engineering.
MFng Is Dispensable for Mouse Pancreas Development and Function