The developmental potential of mouse oocytes matured in serum-free culture conditions.
Studies were undertaken to identify serum-free conditions for the maturation of mouse oocytes in vitro. Oocytes were recovered from the antral follicles of juvenile mice 48 h after injection with gonadotrophin and allowed to resume meiosis in modified Ham's F-10 (mHF-10) medium unsupplemented or supplemented with bovine serum albumin (BSA), fetal calf serum, human pre-ovulatory serum, human follicular fluid or EDTA. They were inseminated 14-16 h later, scored for polar body extrusion after 4-6 h with spermatozoa, and transferred to protein-free mHF-10 for further development. In-vivo matured ova were inseminated and cultured in parallel as controls. Fertilization and development were scored as two cells 24 h after insemination and blastocysts 4 days following insemination respectively. Surprisingly, 41% of oocytes cultured in unsupplemented mHF-10 completed meiosis I, and of those, 50% fertilized; serum supplementation did not improve maturation or fertilization rates. Although the addition of human follicular fluid to the mHF-10 improved meiosis (69%) and fertilization (68% of eggs with polar bodies) to levels comparable with the in-vivo control eggs (79 and 66% respectively), BSA supplementation was equally beneficial. Blastocyst development varied, but within each maturation/fertilization group, the development from in-vitro matured eggs was comparable with embryos from in-vivo matured eggs. In addition, two out of eight 4-cell embryos from oocytes cultured in mHF-10 with BSA and EDTA gave rise to apparently normal pups following transfer to pseudopregnant recipients.(ABSTRACT TRUNCATED AT 250 WORDS)
Serta
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RT
Deaconess/Harvard Surgical Service, Laboratory of Reproductive Biology, Boston, MA 02215, USA.
Michalopoulos
J
J
Seibel
M M
MM
Kiessling
A A
AA
D000818:Animals; D001755:Blastocyst / Q000502:physiology; D002417:Cattle; D016922:Cellular Senescence; D016895:Culture Media, Serum-Free; D046508:Culture Techniques; D010064:Embryo Implantation; D004624:Embryo Transfer; D005314:Embryonic and Fetal Development; D005260:Female; D005306:Fertilization; D006801:Humans; D008540:Meiosis; D051379:Mice; D008815:Mice, Inbred Strains; D009865:Oocytes / Q000502:physiology*
D016895:Culture Media, Serum-Free