Duct, exocrine, and endocrine components of cultured fetal mouse pancreas

Abstract

Twenty to twenty-two days postcoitum mouse fetal pancreas organ bits were cultured on the dermal surface of irradiated pigskin as a substrate. The medium used for long term culture consisted of Eagle's Minimum Essential Medium with the addition of 10% bovine serum, 0.02 U/ml insulin, 0.025 microgram/ml glucagon, 3.63 microgram/ml hydrocortisone, 100 microgram/ml soybean trypsin inhibitor or 10(-8) M atropine. When the medium lacked trypsin inhibitor or atropine but contained the three hormones, the pigskin support began to be destroyed after 2 to 4 wk in culture. Thereafter, the cultured cells could not grow and survive on the digested pigskin. When 10(-6) M atropine was added to the medium, amylase secretion from cultured cells and destruction of pigskin were inhibited completely but pancreas cells could not grow or survive. In contrast, 100 microgram/ml soybean trypsin inhibitor or 10(-8) M atropine permitted cell growth, permitted amylase secretion from the cultured acinar cells, and prevented the destruction of pigskin. Under these conditions pancreas cells migrated or grew or both from the organ bits onto the surface of the pigskin dermis and organoid aggregations formed. Hydrocortisone was needed to permit growth for more than 2 wk. Glucagon and insulin had additive effects. Light and electron microscopic observations indicated the culture of at least five kinds of cells, i.e., duct, acinar, centroacinar, endocrine, and mesenchymal. The majority of cultured cells were duct cells and acinar cells. There were few mesenchymal cells. Mouse pancreas cells were cultured for at least 12 wk by this method.

In vitro Dedifferentiation of Fetal Porcine Pancreatic Tissue prior to Transplantation as Islet-Like Cell Clusters

“…The loss of exocrine morphology and antigenicity and consequent development of the expression of ductal antigens has also been described in a number of in vivo models of adult pancreatic regeneration [Bockman and Merlino, 1992;Bonner-Weir et al, 1993;Gu et al, 1994;Wang et al, 1995]. When explants of mid fetal human pancreas [Lazarow et al, 1973], late fetal rat [Lazarow et al, 1973;McEvoy et al, 1973] and mouse [Hirata et al, 1982] were cultured for 4 days, there was a loss of exocrine cells but no significant report of cell death. Consequently there was a loss of acinar cell mass but a less significant loss of explant mass, a situation similar to our findings where the maintenance of explant mass is probably attributable to exocrine cell dedifferentiation as opposed to cell deletion.…”