In vitro development of pig preantral follicles cultured in a serum-free medium and the effect of angiotensin II

A novel culture system is reported in which pig preantral follicles (< 300 microm in diameter) with an intact thecal cell layer were isolated and cultured in a serum-free medium for up to 30 days. The medium supported follicle culture after isolation, while maintaining both somatic cell and oocyt...

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Vydáno v:Reproduction (Cambridge, England) Ročník 123; číslo 6; s. 807
Hlavní autoři: Shuttleworth, G, Broughton Pipkin, F, Hunter, M G
Médium: Journal Article
Jazyk:angličtina
Vydáno: England 01.06.2002
Témata:
Angiotensin II - pharmacology
Animals
Cell Division
Cell Survival
Culture Media, Serum-Free
Culture Techniques - methods
Estradiol - biosynthesis
Female
Ovarian Follicle - drug effects
Progesterone - biosynthesis
Swine
Testosterone - pharmacology
Time Factors
ISSN:1470-1626
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Shrnutí:A novel culture system is reported in which pig preantral follicles (< 300 microm in diameter) with an intact thecal cell layer were isolated and cultured in a serum-free medium for up to 30 days. The medium supported follicle culture after isolation, while maintaining both somatic cell and oocyte viability. Follicles were cultured in groups (n = 3 per group) on collagen-coated wells for 16 days, during which they retained a three-dimensional structure, maintained oocyte viability and increased in diameter and number of somatic cells. Follicle culture for 30 days resulted in a further increase in number of cells, oocyte viability was maintained, and a significant increase in follicle diameter was observed (P < 0.001), with 29% of follicles forming an antrum. Follicles synthesized measurable quantities of progesterone (168 pg per 100 microl per 48 h; no significant increase with time) and increasing quantities of oestradiol (136 pg per 100 microl per 48 h; P < 0.001 with time). Further supplementation of the medium with 100 micromol testosterone l(-1) at day 28 resulted in a significant increase in oestradiol secretion by both antral (P < 0.01) and preantral follicles (P < 0.05). Culture over 30 days in medium with 10(-10) mol angiotensin II l(-1) and further supplementation at day 28 with 100 micromol testosterone l-1 also increased oestradiol synthesis (P < 0.001). These results show that viable preantral follicles may be cultured for extended periods, and indicate that the possible role of angiotensin II in folliculogenesis and steroidogenesis in early development of pig follicles requires further investigation.
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ISSN:1470-1626
DOI:10.1530/rep.0.1230807