Adrenal medullary chromaffin cell culture systems are extremely useful for the study of excitation-secretion coupling in an in vitro setting. This protocol illustrates the method used to dissect the adrenals and then isolate the medullary region by stripping away the adrenal cortex. The digestion of the medulla into single chromaffin cells is then demonstrated.
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Summary
Abstract
Adrenal medullary chromaffin cell culture systems are extremely useful for the study of excitation-secretion coupling in an in vitro setting. This protocol illustrates the method used to dissect the adrenals and then isolate the medullary region by stripping away the adrenal cortex. The digestion of the medulla into single chromaffin cells is then demonstrated.
Protocol
Abbreviations:
ACCs - Adrenal Chromaffin Cells, E.Soln - Enzyme Solution, E.DMEM - Enriched Dulbecco's Modified Eagle Medium
- Preparation
- Add papain to desired quantity of E. Soln (40 units papain/ 1ml E. Soln.).
- Activate papain with carbogen for about 15 minutes on ice. Oxygenate Locke's buffer on ice as well.
- Dissection
- Use 8-10 week mouse.
- Prior to digestion remove oxygenated Locke's buffer and place on ice.
- Refer to video for dissection procedure.
- Once exci....
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Disclosures
The authors have nothing to disclose.
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