In vitro culture on Matrigel favors the long-term maintenance of rat zona glomerulosa-cell differentiated phenotype
Zona glomerulosa (ZG) cells cultured on plastic within few days dedifferentiate losing their capacity to secrete aldosterone (ALDO) in appreciable amounts. Evidence indicates that extracellular matrix modulates the secretory behavior of adrenocortical cells cultured in vitro. Hence, we compared the...
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Veröffentlicht in: | International journal of molecular medicine 2006-06, Vol.17 (6), p.1101-1110 |
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Sprache: | eng |
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Zusammenfassung: | Zona glomerulosa (ZG) cells cultured on plastic within few days dedifferentiate
losing their capacity to secrete aldosterone (ALDO) in appreciable amounts. Evidence
indicates that extracellular matrix modulates the secretory behavior of adrenocortical
cells cultured in vitro. Hence, we compared the morphology and function of rat
ZG cells grown on plastic and Matrigel basement membrane matrix (herein-after
Matrigel) for up to 12 days. At day 3, no significant differences were observed
between cells cultured on plastic and Matrigel. Starting from day 6, ZG cells
cultured on plastic lost their ultrastructural differentiated features (mitochondria
with tubular cristae, smooth endoplasmic reticulum cisternae and lipid droplets),
exhibiting a fibroblast-like appearance. The mRNA expression of the main steroidogenic
enzymes, as evaluated by real-time polymerase chain reaction, the baseline secretion
of ALDO and other post-pregnenolone hormones, as evaluated by high pressure liquid
chromatography, and the secretory response to ACTH, angiotensin-II and K+, as
evaluated by radioimmunoassay, displayed a time-dependent decrease. Matrigel was
found to maintain unchanged both the ultrastructure and the expresion of steroidogenic
enzymes of ZG cells until day 12 of culture. Baseline and agonist-stimulated steroid-hormone
secretion decreased with the duration of culture on Matrigel, but was always higher
than that of ZG cells grown on plastic. Hence, our study clearly indicates that
the culture on Matrigel favors the maintenance of rat ZG-cell differentiated phenotype,
allowing the conclusion that this technique is suitable for long-term in vitro
investigations. |
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ISSN: | 1107-3756 1791-244X |
DOI: | 10.3892/ijmm.17.6.1101 |