Effects of granulosa coculture on in-vitro oocyte meiotic maturation within a putatively less competent murine model.
A less competent murine in vitro maturation (IVM) model was achieved by shortening the standard duration of in vivo PMSG stimulation from 48 to 24 h and selecting only naked/partially naked GV oocytes from a mixture of large and small follicles. Porcine granulosa coculture enhanced meiotic maturation within such a less competent model (37.3% versus 23.1%, P<0.05), while no significant enhancement was observed with macaque and murine granulosa coculture. Culture of porcine granulosa on extracellular matrix (ECM) gel resulted in a more differentiated morphology, but did not significantly further enhance the beneficial effects it already had on meiotic maturation. Increased concentrations of serum as well as the supplementation of gonadotrophins and follicular fluid within the culture milieu did not enhance IVM under both cell-free and coculture conditions. Porcine granulosa-conditioned medium also enhanced meiotic maturation (36.5% versus 26.7%, P<0.05), which was not diminished upon freeze-thawing (35.8% versus 22.6%, P<0.05). Enhancement of meiotic maturation by porcine granulosa coculture did not however translate to significant improvements in developmental competence, as assessed by in vitro fertilization (IVF) and embryo culture to the blastocyst stage, followed by total cell counts. ECM gel had a detrimental effect on fertilization and developmental competence, even though it had no detrimental effect on meiotic maturation itself.
Department of Obstetrics & Gynaecology, Faculty of Medicine, National University of Singapore, 5 Lower Kent Ridge Road, Singapore 119074.
Boon Chin
BC
Heng
Guo Qing
GQ
Tong
Soon Chye
SC
Ng
D000818:Animals; D001755:Blastocyst / Q000502:physiology; D001769:Blood; D002478:Cells, Cultured; D018920:Coculture Techniques; D017077:Culture Media, Conditioned; D046508:Culture Techniques; D005314:Embryonic and Fetal Development; D005109:Extracellular Matrix; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D015571:Follicular Fluid; D006064:Gonadotropins, Equine / Q000008:administration & dosage; D006107:Granulosa Cells / Q000502:physiology*; D008252:Macaca fascicularis; D008540:Meiosis; D051379:Mice; D023421:Models, Animal; D009865:Oocytes / Q000166:cytology* / Q000254:growth & development; D013552:Swine
D017077:Culture Media, Conditioned; D006064:Gonadotropins, Equine