Analysis and optimization of nutritional set-up for murine pancreatic acinar cells.
BACKGROUND
Pancreatic acinar cell cultivation poses a serious problem due to limitations in the in vitro survival time despite variations of dissociation protocols, culture media and nutrient supplements.


OBJECTIVE
To establish a long term culture of murine pancreatic acinar cells which retain their viability, monolayer formation and responsiveness to secretagogues. In order to investigate the mechanism of the short-life of acinar cells studied in vitro, we studied their survival under the influence of different supplements on nutrient media.


METHODS
Dissociated pancreatic acini were prepared from BALB/c mice pancreata by collagenase digestion supplemented with bovine serum albumin fraction V and soybean trypsin inhibitor. A nutrient set-up was designed for their long term survival in vitro.


RESULTS
It was observed that mouse pancreatic acinar cells dissociated in presence of bovine serum albumin fraction V and soybean trypsin inhibitor result in 95% viability. Further cultivation of these acinar cells in Waymouth's MB 752/1 medium supplemented with 10% fetal calf serum (v/v), soybean trypsin inhibitor, bovine serum albumin, dexamethasone, and epidermal growth factor results in their survival for more than 6 days in culture with 85% viability, retention of the secretagogue responsiveness and formation of a monolayer without any extracellular matrix coating.


CONCLUSIONS
Our study clearly demonstrates that the addition of soybean trypsin inhibitor to culture medium reduces zymogen granule fragility and acinar cell death, thus increasing their viability for sufficiently long periods. The present study offers an excellent, in vitro model for the investigation of exocrine dysfunction in response to acinar cell injury.
Kurup
Savita
S
National Center for Cell Science, University of Pune Campus, Ganeshkhind, Pune, India.
Bhonde
Ramesh R
RR
D000681:Amylases / Q000378:metabolism; D000818:Animals; D002469:Cell Separation; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D003470:Culture Media / Q000032:analysis; D005260:Female; D006023:Glycoproteins / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009751:Nutritional Requirements; D010179:Pancreas / Q000166:cytology* / Q000201:enzymology / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000378:metabolism
D003470:Culture Media; D006023:Glycoproteins; D012710:Serum Albumin, Bovine; D000681:Amylases