Related Experiment Videos
Amylase secretion from isolated pure acinar cells.
Biochemical and Biophysical Research Communications
|May 31, 1985
Summary
Researchers isolated pure rat pancreatic acinar cells using counterflow sedimentation filtration technique (CSFT). These cells responded to cholecystokinin (CCK8) and other peptides, enabling studies on pancreatic secretion.
Area of Science:
- Biochemistry
- Cell Biology
- Physiology
Background:
- Pancreatic acinar cells are crucial for digestive enzyme secretion.
- Previous methods for isolating pure acinar cells were often inefficient or resulted in compromised cell integrity.
- Understanding acinar cell function requires highly purified, viable cell preparations.
Purpose of the Study:
- To establish a reliable method for isolating pure pancreatic acinar cells from rats.
- To assess the functional integrity and responsiveness of these isolated cells to various secretagogues.
- To investigate the effects of specific peptides on amylase secretion from purified acinar cells.
Main Methods:
- Isolation of rat pancreatic acinar cells utilizing counterflow sedimentation filtration technique (CSFT).
- Assessment of cell purity by microscopic examination, ensuring absence of endocrine and duct cells.
- Measurement of amylase secretion in response to octapeptide of cholecystokinin (CCK8), insulin, glucagon, and vasoactive intestinal polypeptide (VIP).
- Evaluation of cell integrity through lactic dehydrogenase (LDH) leakage assays.
Main Results:
- CSFT yielded highly purified acinar cells with minimal red blood cell contamination and no endocrine or duct cells.
- Octapeptide of cholecystokinin (CCK8) significantly stimulated amylase secretion.
- Insulin and vasoactive intestinal polypeptide (VIP) potentiated CCK8-induced amylase secretion.
- Synthetic glucagon inhibited basal and CCK8-stimulated amylase secretion, while non-synthetic glucagon stimulated secretion.
- No significant lactic dehydrogenase (LDH) leakage was observed, indicating excellent cell viability.
Conclusions:
- Counterflow sedimentation filtration technique (CSFT) provides a superior method for obtaining pure, functionally intact rat pancreatic acinar cells.
- Isolated acinar cells are suitable for studying pancreatic secretion mechanisms and the modulatory effects of various peptides.
- This preparation method facilitates research into pancreatic exocrine function and related signaling pathways.