Linked Life Data

10906754

Source:http://linkedlifedata.com/resource/pubmed/id/10906754

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
2000-9-29
pubmed:abstractText
Adrenal chromaffin cells have been successfully used to attenuate chronic pain when transplanted near the spinal cord, but primary cells are neither homogeneous nor practical for routine use in human therapy. Conditional immortalization with the temperature-sensitive allele of the large T antigen (tsTag) and creation of stable chromaffin cell lines would advance our understanding of both the use and limits of cell lines that contain this immortalization gene for such therapies. Cultures of embryonic day 17 rat adrenal and neonatal bovine adrenal cells were immortalized with the temperature-sensitive allele of SV40 tsTag and chromaffin cell lines established. The rat chromaffin line, RAD5.2, and the bovine chromaffin cell line, BADA.20, both expressed immunoreactivities (ir) for all the catecholamine enzymes: tyrosine hydroxylase (TH), the first enzyme in the synthetic pathway for catecholamines, dopa-beta-hydroxylase (DbetaH), and phenylethanolamine-N-methyltransferase (PNMT). At permissive temperature (33 degrees C), these chromaffin cells are proliferative, have a typical rounded chromaffinlike morphology, and contain detectable TH-, DbetaH-, and PNMT-ir. At nonpermissive temperature (39 degrees C), these cells stop proliferating, decrease Tag expression, and change the expression of TH-, DbetaH-, and PNMT-ir in vitro, suggesting increased differentiation at nonpermissive temperature. The chromaffin cell lines also express immunoreactivity for the opioid met-enkephalin (ENK) at permissive and nonpermissive temperatures. The expression of TH-ir in the bovine chromaffin cells is upregulated by the addition of dexamethasone (DEX) or forskolin during differentiation; TH-ir is not affected by the addition of DEX or forskolin in the rat chromaffin cells. The addition of forskolin during differentiation upregulates the expression of DbetaH-ir in the rat chromaffin cells. PNMT-ir is not affected by differentiation or agents in either cell line. However, catecholamine synthesis was not detectable by high-performance liquid chromatography, suggesting incomplete differentiation under current conditions, or influence by continued low levels of Tag expression. Both cell lines have been carried over many passages in vitro for more than 3 years and were repeatedly frozen and thawed. These data describe an initial step in the conditional immortalization of chromaffin cells that can maintain the phenotype of primary chromaffin cells in vitro over long periods. The use of such chromaffin cell lines that are able to deliver neuroactive molecules offers a novel approach to pain management.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jul
pubmed:issn
0730-2312
pubmed:author
pubmed:copyrightInfo
Copyright 2000 Wiley-Liss, Inc.
pubmed:issnType
Print
pubmed:day
19
pubmed:volume
79
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
38-57
pubmed:dateRevised
2011-11-17
pubmed:meshHeading
pubmed-meshheading:10906754-Adrenal Glands, pubmed-meshheading:10906754-Animals, pubmed-meshheading:10906754-Catecholamines, pubmed-meshheading:10906754-Cattle, pubmed-meshheading:10906754-Cell Differentiation, pubmed-meshheading:10906754-Cell Line, Transformed, pubmed-meshheading:10906754-Chromaffin Cells, pubmed-meshheading:10906754-Chromatography, High Pressure Liquid, pubmed-meshheading:10906754-Dexamethasone, pubmed-meshheading:10906754-Dopamine beta-Hydroxylase, pubmed-meshheading:10906754-Enkephalin, Methionine, pubmed-meshheading:10906754-Female, pubmed-meshheading:10906754-Forskolin, pubmed-meshheading:10906754-Hot Temperature, pubmed-meshheading:10906754-Humans, pubmed-meshheading:10906754-Immunohistochemistry, pubmed-meshheading:10906754-Pain, pubmed-meshheading:10906754-Pain Management, pubmed-meshheading:10906754-Peripheral Nerve Injuries, pubmed-meshheading:10906754-Phenylethanolamine N-Methyltransferase, pubmed-meshheading:10906754-Rats, pubmed-meshheading:10906754-Spinal Cord Injuries, pubmed-meshheading:10906754-Tyrosine 3-Monooxygenase
pubmed:year
2000
pubmed:articleTitle
Generation and initial characterization of conditionally immortalized chromaffin cells.
pubmed:affiliation
The Miami Project To Cure Paralysis, University of Miami School of Medicine, Miami, Florida 33136, USA. meaton@miami.edu
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't