بررسي اثر همزمان ماتريكس برون سلولي و همكشتي پوياي سلو لهاي پانكراس جنين موش بر تمايز سلولهاي مولد انسولين از سلولهاي بنيادي جنيني موش

Simultaneous Effect of Extracellular Matrix and Embryonic Pancreas Dynamic Co-culture on Differentiation of Insulin-Producing Cells From Mouse Embryonic Stem Ceils

Purpose: This study initiated to examine supportive effect of embryonic pancreas secreted agents as well as collagen, gelatin and matrigel as extracellular matrix on which cells are differentiated. Materials and Methods: Protocol started with mouse embryonic stem cell line (Royan B1). At the final stage, differentiating cells were replated onto different matrices and filter inserts explant-containing embryonic pancreas were transferrd to above of them. Expression of pancreas-specific or -enriched genes was examined by RT-PCR; Immunostainig of insulin molecule was performed; de novo expression of insulin was also verified using anti C-peptide antibody. Insulin- and C-peptide- positive cells were also counted. Results: Pancreas-specific genes were expressed in cells which were co-cultured and differentiated on collagen or gelatin. These cells were immuno-reactive with insulin and C-peptide antibodies. The maximum percentages of insulin- and C-peptide -positive cells obtained in cells which were co-cultured and differentiated on collagen (3.99 ± 0.23 % and 1.94 ± 0.06 %, respectively). Conclusion: These results suggest that extracellular matrix (collagen and its denatured form, gelatin) has an important implication on differentiation of mouse embryonic stem cells to insulin-producing cell. In addition, embryonic pancreatic cells release molecules that promote the differentiation.