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Titolo:
Generation of stable cell lines by spontaneous immortalization of primary cultures of porcine granulosa cells
Autore:
Gillio-Meina, C; Swan, CL; Crellin, NK; Stocco, DM; Chedrese, PJ;
Indirizzi:
Univ Saskatchewan, Dept Obstet Gynecol & Reprod Sci, Saskatoon, SK, CanadaUniv Saskatchewan Saskatoon SK Canada Reprod Sci, Saskatoon, SK, Canada Univ Saskatchewan, Toxicol Ctr, Saskatoon, SK S7N 0W0, Canada Univ Saskatchewan Saskatoon SK Canada S7N 0W0 skatoon, SK S7N 0W0, Canada Texas Tech Univ, Hlth Sci Ctr, Dept Cell Biol & Biochem, Lubbock, TX 79430USA Texas Tech Univ Lubbock TX USA 79430 Biol & Biochem, Lubbock, TX 79430USA
Titolo Testata:
MOLECULAR REPRODUCTION AND DEVELOPMENT
fascicolo: 4, volume: 57, anno: 2000,
pagine: 366 - 374
SICI:
1040-452X(200012)57:4<366:GOSCLB>2.0.ZU;2-W
Fonte:
ISI
Lingua:
ENG
Soggetto:
3-BETA-HYDROXY-5-ENE STEROID DEHYDROGENASE; PROTEIN STAR GENE; PROGESTERONE SYNTHESIS; GROWTH-FACTOR; EXPRESSION; KINASE; TRANSFORMATION; STEROIDOGENESIS; ESTABLISHMENT; HORMONE;
Keywords:
immortalization; granulosa cells; steroidogenesis; StAR; P450scc; 3 beta-HSD; P450arom;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
48
Recensione:
Indirizzi per estratti:
Indirizzo: Chedrese, PJ Royal Univ Hosp, Dept Obstet Gynecol & Reprod Sci, 103 Hosp Dr, Saskatoon,SK S7N 0W8, Canada Royal Univ Hosp 103 Hosp Dr Saskatoon SK Canada S7N 0W8 anada
Citazione:
C. Gillio-Meina et al., "Generation of stable cell lines by spontaneous immortalization of primary cultures of porcine granulosa cells", MOL REPROD, 57(4), 2000, pp. 366-374

Abstract

We report the generation of stable cell lines obtained by spontaneous immortalization of primary cultures of porcine granulosa cells. Three hundred stable cell lines were obtained from three independent immortalization trials. Two of these cell lines retained the steroidogenic capabilities characteristic of granulosa cells, such as de novo synthesis of progesterone and conversion of androstenedione into estradiol-17 beta. All the stable cell lines expressed the P450arom and 3 beta HSD genes, confirming their granulosa origin. Moreover, the steroidogenic stable granulosa cells also expressed StAR and P450scc genes. Stable cells were developed in cultures using Medium199 supplemented with 5% newborn calf serum (NBCS). The surviving cells overcame the senescent phase and entered a stage of continuous growth for over one hundred generations. No stable colonies were obtained from cultures grown in MEM or DMEM or media supplemented with 10% NBCS or 5 and 10% fetal calf serum (FCS). Medium 199 is a formulation richer in nutrients compared to MEM or DM EM and the cell growth capability of NBCS is lower than that of FCS, probably due to deficiency of growth factors. We speculate that spontaneous immortalization of granulosa cells may be facilitated by using a rich culture formulation supplemented with low concentrations of serum deficient in growth factors. We have validated the stable cell lines for studyingthe effect of hormonal steroids on granulosa cell steroidogenesis and the expression of the steroidogenic genes. Therefore, we believe that they are useful models to study the molecular mechanism involved in granulosa cell differentiation and steroidogenesis. (C) 2000 Wiley-Liss, Inc.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 27/09/20 alle ore 22:52:23