Culture of porcine fetal pancreatic neurons

Objectives: Pancreatic neurons have not been cultured commonly. Cultured neurons can be continuously observed, and their external environment is easy to be controlled. We report here a simple method for separating and cultivating neuronal cells from pancreas. Methods: Pancreata of fetal swine were digested with collagenase. Clusters were collected with a sieve and digested with trypsin. Digested clusters were collected and cultured in Dulbecco modified Eagle medium containing serum and basic fibroblast growth factor. Cultured cells were investigated morphologically. Results: Cultured cells formed spiderweblike colonies. These cells were distinguished into Schwann cells and 2 types of neurons. The neurons were positive on immunocytochemical staining with anti-growth-associated protein-43 and cytochemical staining for cholinesterase. One type of neuron was located in the central cluster and had very long processes extending radially. The other type of neuron was sparsely scattered, had long processes, and was connected to other neurons. The neurotransmitter of these neurons was concluded to be acetylcholine. Conclusions: Using our method, neuronal cells were readily cultured from pancreatic tissue. These cells will be useful in elucidating the physiology and pharmacology of pancreatic neurons.