Primary culture of pancreatic (human) acinar cells.
The acinar cell culture plays a very important role in research of pancreatic pathophysiology. The aim of this study was to establish a long-term culture of human (foetal) pancreatic acinar cells in standardized nutrient media with supplements. Acinar cells were prepared from pancreatic tissues obtained from aborted foetus (> or =35 weeks) with no prior pancreatic complications by collagenase digestion and cultured using different media and supplements. The purity and phenotype of acinar cells was confirmed by various staining techniques and FACS. The acinar cell proliferation was determined at different time intervals by Bromo-deoxyuridine (BrdU) incorporation, and metabolic enzyme activity was analysed. The acini could be cultured and maintained in Ham's F-12 K/M199 media in the presence of 5% BSA, 0.1 mg/ml STI, 10 ng/ml EGF, and 10% FCS with the same morphological appearance as that of freshly prepared for 12 days with maximum viability of 80-85% and formation of monolayer without extracellular matrix. A significant BrdU incorporation of acinar cells in primary culture was observed which was maximum (105%) at day four. Higher amylase and lipase activity was seen in freshly isolated acinar cells which decreased with time of the culture. The established human pancreatic acinar cell culture may act as an excellent model to study exocrine dysfunction or pancreatitis in response to acinar cell injury.
Singh
Lipi
L
Department of General Surgery, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
Bakshi
Dapinder K
DK
Vasishta
Rakesh Kumar
RK
Arora
Sunil Kumar
SK
Majumdar
Siddarth
S
Wig
Jai Dev
JD
D000681:Amylases / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D002469:Cell Separation; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D006801:Humans; D008049:Lipase / Q000378:metabolism; D010179:Pancreas / Q000166:cytology* / Q000201:enzymology / Q000502:physiology*
D008049:Lipase; D000681:Amylases