Culture of porcine fetal pancreatic neurons.
OBJECTIVE
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 antigrowth-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.
Kuroki
Masahiko
M
Department of Anatomy and Neurobiology, National Defense Medical College, Tokorozawa, Saitama, Japan. kuroki@ri.ndmc.ac.jp
Sawasaki
Yoshio
Y
Kuroki
Kazuyo
K
Takeuchi
Kyoko
K
Yoshioka
Masahiko
M
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D002802:Cholinesterases / Q000378:metabolism; D005333:Fetus / Q000166:cytology / Q000294:innervation / Q000378:metabolism; D019922:GAP-43 Protein / Q000378:metabolism; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D007150:Immunohistochemistry; D004676:Myelin Basic Protein / Q000378:metabolism; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D010179:Pancreas / Q000166:cytology* / Q000294:innervation* / Q000378:metabolism; D013552:Swine
D019922:GAP-43 Protein; D005904:Glial Fibrillary Acidic Protein; D004676:Myelin Basic Protein; D002802:Cholinesterases