Automated Method for Isolation of Adrenal Medullary Chromaffin Cells from Neonatal Porcine Glands

An automated method for the isolation of neonatal porcine adrenal chromaffin cells is described. Adrenal chromaffin cells are potentially useful for therapeutic transplantation, but current isolation methodology suffers from labor intensiveness and variability in yield and viability due to imprecisi...

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Bibliographic Details
Published in:Cell transplantation 2001-01, Vol.10 (8), p.689-696
Main Authors: Vizzardelli, Caterina, Potter, Elizabeth D., Berney, Thierry, Pileggi, Antonello, Inverardi, Luca, Ricordi, Camillo, Sagen, Jacqueline
Format: Article
Language:English
Subjects:
Adrenal Medulla - cytology
Animals
Animals, Newborn
Automation
Cell Separation - methods
Cell Survival
Cells, Cultured
Chromaffin Cells - cytology
Chromaffin Cells - metabolism
Epinephrine - biosynthesis
Epinephrine - secretion
Norepinephrine - biosynthesis
Norepinephrine - secretion
Swine
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Summary:An automated method for the isolation of neonatal porcine adrenal chromaffin cells is described. Adrenal chromaffin cells are potentially useful for therapeutic transplantation, but current isolation methodology suffers from labor intensiveness and variability in yield and viability due to imprecision of manual techniques, enzyme purity, and gland age and species. The described approach utilizes an adaptation of an automated procedure previously described for isolation of pancreatic islets. Results from neonatal porcine adrenal glands revealed consistent cell yields with high (approximately 99%) viability. Catecholamine assays showed that the resultant cultures continue to produce and secrete norepinephrine and epinephrine. Immunocytochemical analysis indicated that the majority of cells in the preparation are chromaffin cells and adrenal cortical cells. The procedure meets the following requirements: 1) minimal traumatic action on the adrenal chromaffin cells, 2) continuous digestion in which the adrenal cells that are progressively liberated can be saved from further mechanical action, 3) minimal human intervention in the digestion process, and 4) high yield and viability of the isolated adrenal chromaffin cells.
ISSN:0963-6897
1555-3892
DOI:10.3727/000000001783986288