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Indian J Exp Biol ; 2015 Feb; 53(2): 75-81
Article Dans Anglais | IMSEAR | ID: sea-158379

Résumé

The bidirectional communication between oocytes and granulosa cells are mediated by several factors via a local feedback loop(s). The current model was carried out to study the spatial mutual interaction of porcine denuded oocytes and granulosa cells either in direct contact (juxtacrine) or paracrine co-culture using transwell system. Transwell 0.4 µm polyester membrane inserts were used to permit oocytes-granulosa cells paracrine communication with a distance of 2 mm between them in co-culture. Oocytes were cultured with granulosa cells in a defined basic maturation medium for 44 h. In results, oocyte secreted factors (OSFs; GDF9 and BMP15) temporal expression showed progressive decrement by the end of culture in case of direct contact with granulosa cells while it was increased progressively in the paracrine co-culture groups. However, oocytes that were cultured in direct contact showed a significant increase in blastocyst development after parthenogenetic activation than the paracrine co-cultured ones (20% vs. 11.5%, respectively). By the end of culture, granulosa cell count in direct contact showed a significant decrease than the indirect co-culture group (1.2 × 105 cell/mL vs. 2.1 × 105 cell/mL, respectively). Steroids (P4 and E2) and steriodogenesis enzymes mRNA levels showed significant temporal alterations either after 22 h and 44 h of IVM in both juxtacrine and paracrine co-culture systems (P ≤ 0.05). CX43 was much more highly expressed in the granulosa of the direct contact group than the indirect co-culture group. These results indicate the difference in mutual communication between oocytes and granulosa cells that were cocultured either in direct contact (juxtacrine) or with a short distance (paracrine) and propose a new paradigm to study different ovarian follicular cells interaction.


Sujets)
/génétique , /métabolisme , Animaux , Aromatase/génétique , Aromatase/métabolisme , Protéine morphogénétique osseuse de type 15/génétique , Protéine morphogénétique osseuse de type 15/métabolisme , Communication cellulaire , Cellules cultivées , Techniques de coculture/méthodes , Connexine 43/génétique , Connexine 43/métabolisme , Oestradiol/métabolisme , Femelle , Jonctions communicantes/métabolisme , Expression des gènes , Cellules de la granulosa/cytologie , Cellules de la granulosa/métabolisme , Facteur-9 de croissance et de différenciation/génétique , Facteur-9 de croissance et de différenciation/métabolisme , Ovocytes/cytologie , Ovocytes/métabolisme , Communication paracrine , Progestérone/métabolisme , RT-PCR , Suidae
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