Effects of follicular fluid or progesterone on in vitro maturation of equine oocytes before intracytoplasmic sperm injection with non-sorted and sex-sorted spermatozoa.
In Expt 1, compact cumulus oocyte complexes (COCs) were matured in: (i) control medium (Hepes-buffered TCM-199 with 10% oestrous cow serum (OCS) + oestradiol, LH and FSH); (ii) Hepes-buffered TCM-199 with 20% follicular fluid; or (iii) control medium containing 250 ng progesterone ml(-1). Mature oocytes were collected by transvaginal aspiration as a positive control for the in vitro maturation (IVM) treatments. Oocytes were fertilized by ICSI and cultured in Menezo's B2 + 5% fetal calf serum (FCS). There were no significant differences among IVM treatments. In Expt 2, oocytes with expanded COCs were matured in Hepes-buffered TCM-199 with 10% OCS, oestradiol, LH and FSH with different concentrations of progesterone (0, 50, 250 and 1250 ng ml(-1)). Oocytes were fertilized by ICSI and cultured in a chemically defined medium. The medium containing 1250 ng progesterone ml(-1) resulted in fewer oocytes with a visible first polar body after maturation (P < 0.05), whereas the media containing 0 and 50 ng progesterone ml(-1) resulted in higher development rates to seven- to eight-cell embryos (P < 0.05), compared with media containing 250 or 1250 ng progesterone ml(-1). Six of the resulting morulae were transferred to recipient mares. In addition, oocytes (n=32) from Expt 2 were injected with sex-sorted spermatozoa, obtained by separating X- and Y-bearing spermatozoa with a Cytomation MoFlo flow cytometer/cell sorter. Two embryos resulting from ICSI with X-bearing spermatozoa were transferred to the oviduct of a recipient mare. No pregnancies were established after transfer of embryos in these experiments.
Schmid
R L
RL
Animal Reproduction and Biotechnology Laboratory, Colorado State University, Fort Collins, CO 80523, USA.
Kato
H
H
Herickhoff
L A
LA
Schenk
J L
JL
McCue
P M
PM
Chung
Y G
YG
Squires
E L
EL
D000818:Animals; D004624:Embryo Transfer / Q000662:veterinary; D005260:Female; D006736:Horses / Q000502:physiology*; D008297:Male; D009865:Oocytes / Q000502:physiology*; D006080:Ovarian Follicle; D011247:Pregnancy; D011374:Progesterone / Q000494:pharmacology*; D012743:Sex Preselection / Q000662:veterinary*; D020554:Sperm Injections, Intracytoplasmic / Q000662:veterinary*; D013094:Spermatozoa / Q000502:physiology; D014960:X Chromosome
D011374:Progesterone