Effect of Different Media and Protein Source on Equine Gametes: Potential Impact During In Vitro Fertilization.
Equine in vitro fertilization (IVF) is still inconsistent. In the present work, we studied how modified Whitten's (MW) medium and Tissue Culture Medium 199 (TCM) added with Foetal Bovine Serum (FBS; 10% v/v) or Bovine Serum Albumin (BSA; 7 mg/ml) affected equine gametes to subsequently run IVF trials. Compact (Cp) and expanded (Ex) cumuli equine oocytes were matured and placed in TCM or MW supplemented with BSA or FBS for 18-20 h (no sperm added). In Ex oocytes, TCM-199 added with FBS or BSA resulted in higher metaphase II (MII) rates (75.7% and 62.7%, respectively) than MW added with BSA (54%) or FBS (52.2%; p < 0.05); this was not observed for Cp oocytes. Equine sperm were capacitated in the same media at 10 × 10(6) sperm/ml for 4 h at 37°C; total motility and protein tyrosine phosphorylation (PY) were evaluated. While motility remained unchanged, TCM or MW added with FBS enhanced the number of sperm showing PY-stained tails (25 ± 4.8% and 31 ± 6.6%; mean ± SEM, respectively) over BSA supplemented media (3 ± 1.2% and 11.7 ± 1.1%) for TCM and MW (p < 0.05). In view of the previous results, sperm were capacitated in TCM + FBS and MW + BSA (control); IVF trials were run in the same media supplemented with 200 ng/ml of progesterone, but no fertilization occurred. Our results show that TCM + FBS enhances Ex equine oocyte's meiotic competence over MW + BSA and TCM or MW added with FBS successfully induce equine PY over media supplemented with BSA.
CECA/ICETA - Animal Sciences Centre, University of Porto, Vairão, Portugal.|ICBAS - Abel Salazar Biomedical Institute, University of Porto, Porto, Portugal.
L
http://orcid.org/0000-0001-5568-548X
L
González-Fernández
ICBAS - Abel Salazar Biomedical Institute, University of Porto, Porto, Portugal.
S
S
Macedo
ICBAS - Abel Salazar Biomedical Institute, University of Porto, Porto, Portugal.
J S
JS
Lopes
CECA/ICETA - Animal Sciences Centre, University of Porto, Vairão, Portugal.|ICBAS - Abel Salazar Biomedical Institute, University of Porto, Porto, Portugal.
A
A
Rocha
CECA/ICETA - Animal Sciences Centre, University of Porto, Vairão, Portugal.|ICBAS - Abel Salazar Biomedical Institute, University of Porto, Porto, Portugal.
B
http://orcid.org/0000-0001-5331-9793
B
Macías-García
D000818:Animals; D003470:Culture Media / Q000494:pharmacology*; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary*; D006736:Horses; D008297:Male; D009865:Oocytes / Q000187:drug effects*; D011506:Proteins / Q000494:pharmacology*; D013075:Sperm Capacitation / Q000187:drug effects; D013094:Spermatozoa / Q000187:drug effects*
D003470:Culture Media; D011506:Proteins