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dc.contributor.authorWycherley, G
dc.contributor.authorDowney, D
dc.contributor.authorKane, M T
dc.contributor.authorHynes, A C
dc.date.accessioned2018-08-24T08:26:41Z
dc.date.available2018-08-24T08:26:41Z
dc.date.issued2004-06-01
dc.identifier.citationWycherley, G; Downey, D; Kane, M T; Hynes, A C (2004). A novel follicle culture system markedly increases follicle volume, cell number and oestradiol secretion. Reproduction 127 (6), 669-677
dc.identifier.issn1470-1626,1741-7899
dc.identifier.urihttp://hdl.handle.net/10379/9993
dc.description.abstractThis study reports a novel, simple method for culture of mouse follicles which results in follicles with cell numbers similar to in vivo fully grown follicles. Using this method, follicles (1180-240 mum in diameter) were cultured in a 100 mul inverted drop of medium without oil and compared with culture in upright drops with and without a mineral oil overlay. Follicles, isolated from C57BL/6 x CBA/ca crossbred and MF1 inbred mice, were cultured individually at 37degreesC in 96-well round-bottomed suspension cell tissue culture plates for 6 days. Follicles grown in the inverted drop culture system reached a markedly higher final diameter (means+/- S.E.M.; 471 +/- 6.0 mum) as compared with the upright with oil (363 +/- 2.7 mum) and without oil (358 +/- 4.0) systems. There was no significant effect of mouse strain on follicle diameter. Follicular secretion of oestradiol and lactate into the medium was measured on days 2, 4 and 6 of culture. Secretion of oestradiol per follicle on day 6 was 2.49 +/- 0.45 ng in the inverted and 0.90 +/- 0.17 ng in the upright without oil system (P < 0.001). Follicular secretion of lactate on a per unit of follicle volume basis remained constant in the inverted system over days 2, 4 and 6 and was less (P < 0.001) than secretion in both the upright with and without oil systems. Follicle cell proliferation was markedly increased in the inverted as compared with the upright with oil system; the increases in cell numbers were significant on day 3 (P < 0.01) and on all subsequent days (P < 0.001). These results are discussed in relation to the supply of oxygen to the follicle in culture.
dc.publisherBioScientifica
dc.relation.ispartofReproduction
dc.subjectmouse ovarian-follicles
dc.subjectdevelopment in-vitro
dc.subjectdevelopmental competence
dc.subjectprimordial follicles
dc.subjectlactate production
dc.subjectoxygen
dc.subjectoocytes
dc.subjectgrowth
dc.subjectmetabolism
dc.subjectkinetics
dc.titleA novel follicle culture system markedly increases follicle volume, cell number and oestradiol secretion
dc.typeArticle
dc.identifier.doi10.1530/rep.1.00040
dc.local.publishedsourcehttp://www.reproduction-online.org/content/127/6/669.full.pdf
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