different coating conditions regular control fibronectin and extra cellular matrix ecm coated plates all cells were cultured in sm95 m7 9 1 for 3 day for each passage at passage 6 cells were cultured in mm1 and mm2 medium for 3 days respectively after culture in mm2 cells were collected for isolation of total rna 2 expression of epha4 in adult pancreas and primary pancreatic cell cultures epha4 plays important role of sorting and cell specification during development of adult tissues to identify the expression of epha4 in adult pancreas and cell culture immunofluorenscence and icc studies of adult pancreas and primary pancreatic cells were performed as shown in fig2 a epha4 positive cells were restricted to some of islet cells and no positive staining was found in acinar cells expression of epha4 was also found in a subgroup of cultured pancreatic cells fig2 c 3 comparison of expression of insulin in the epha4 and epha4 u2212 cells expression of insulin in epha4 sorted cells was evaluated by real time pcr the mrna levels of insulin in epha4 positive cells were 9 112 times higher than the epha4 negative cells fig3 a and b the high expression of insulin maintained the epha4 positive cells through sequential p2 p6 fig3 table 1 since epha4 positive cells only represent about 5 of the original cell population u03b2 cell lineage cells were greatly enriched by epha4 sorting we compared the expression of pdx 1 in both epha4 positive and epha4 negative cells during different cell passages by real time pcr as shown in fig4 and table 2 epha4 positive cells have 6 10 times higher mrna level of pdx 1 than epha4 negative.