Abstract
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.
| Original language | English |
|---|---|
| Pages (from-to) | 689-696 |
| Number of pages | 8 |
| Journal | Cell Transplantation |
| Volume | 10 |
| Issue number | 8 |
| State | Published - Dec 1 2001 |
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Keywords
- Adrenal gland
- Automated methodology
- Catecholamines
- Chromaffin cells
- Isolation
- Neural transplantation
ASJC Scopus subject areas
- Cell Biology
- Transplantation
Cite this
Automated method for isolation of adrenal medullary chromaffin cells from neonatal porcine glands. / Vizzardelli, Caterina; Potter, Elizabeth D.; Berney, Thierry; Pileggi, Antonello; Inverardi, Luca A; Ricordi, Camillo; Sagen, Jacqueline.
In: Cell Transplantation, Vol. 10, No. 8, 01.12.2001, p. 689-696.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Automated method for isolation of adrenal medullary chromaffin cells from neonatal porcine glands
AU - Vizzardelli, Caterina
AU - Potter, Elizabeth D.
AU - Berney, Thierry
AU - Pileggi, Antonello
AU - Inverardi, Luca A
AU - Ricordi, Camillo
AU - Sagen, Jacqueline
PY - 2001/12/1
Y1 - 2001/12/1
N2 - 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.
AB - 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.
KW - Adrenal gland
KW - Automated methodology
KW - Catecholamines
KW - Chromaffin cells
KW - Isolation
KW - Neural transplantation
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UR - http://www.scopus.com/inward/citedby.url?scp=0035691246&partnerID=8YFLogxK
M3 - Article
C2 - 11814111
AN - SCOPUS:0035691246
VL - 10
SP - 689
EP - 696
JO - Cell Transplantation
JF - Cell Transplantation
SN - 0963-6897
IS - 8
ER -