Primary Culture of Pancreatic (Human) Acinar Cells

Abstract

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.

Chronic Nicotine Exposure In Vivo and In Vitro Inhibits Vitamin B1 (Thiamin) Uptake by Pancreatic Acinar Cells

“…The viability was also assessed on the day of use of the cultured PAC for transport investigations using the trypan blue exclusion method and found to be > 80%. Finally, morphology of the human acinar cells was similar to that described previously [ 34 ].…”

“…Cultured human primary PAC were used for in vitro studies. The human PAC were isolated and maintained under culture conditions [ 34 ] and were used after 3 days in culture (4 days from isolation). This time period was shown to have minimal effect on cellular morphology and no effect on cell differentiation [ 34 ].…”

“…The human PAC were isolated and maintained under culture conditions [ 34 ] and were used after 3 days in culture (4 days from isolation). This time period was shown to have minimal effect on cellular morphology and no effect on cell differentiation [ 34 ]. Collectively, the results show that nicotine inhibits thiamin uptake by PAC and that this inhibition is mediated, at least in part, at the level of transcription of the thiamin transporter genes.…”

Change in the Characteristics of Ca2+ Signaling in Pancreatic Acinar Cells in Culture

“…In this case, the primary culture of acinar cells from pig pancreas maintains the same morphological characteristics and great viability; but, on the other hand, the enzyme activity decreases after several days [14]. In primary cultures of human (fetal) acinar cells similar results have been observed, depicting a reduction of amylase and lipase activity [15].…”

Mitochondrial dysfunction and apoptosis of acinar cells in chronic pancreatitis

“…22,23 In brief, 1 × 10 6 cells/2 ml 199 M/ well were cultured in six-well plates with 50 nM 3,3′-dihexyloxacarbocyanine iodide (DiOC 6 (3)), and a fl uorochrome dye was added for 30 min. [24][25][26] The cells were then washed with sterile PBS to remove excess fl uorochrome, and the concentration of retained DiOC 6 (3) was read by using fl ow cytometry with excitation at 488 nm and emission at 500 nm.…”