Gamete co-incubation duration modulates blastocyst developmental competence in buffalo through regulation of growth factors, steroidogenesis and oxidative stress pathways.

Authors

Garbhit Sharma, Anand Kumar Pandey, Hitesh Jangra, Sandeep Kumar, Pradeep Saini, Rajesh Kumar, Divya Choudhary

Published in

Molecular biology reports. Volume 53. Issue 1. Jul 23, 2026. Epub Jul 23, 2026.

Abstract

Gamete (sperm-oocyte) co-incubation duration during in vitro fertilization (IVF) is a key determinant of embryo developmental competence in buffalo. This study evaluated the effect of gamete co-incubation duration (2-h intervals from 2 to 24 h) on embryo development and blastocyst competence.
A total of 3680 cumulus-oocyte complexes (COCs) were recovered from 1163 buffalo ovaries. Grade A and B COCs (n = 2390) were matured in vitro, and 2324 matured oocytes were subjected to IVF using different gamete co-incubation durations, followed by culture of 2012 presumptive embryos for 7 days. Cleavage, blastocyst development, and developmental competence were assessed using RT-qPCR.
Cleavage rate was significantly affected (P < 0.05), increasing from 65.2% at 2 h to 100% at 8 h and remaining comparable between 10 and 24 h (81.7-97.1%). Blastocyst development increased progressively, reaching the highest rates at 16-20 h (53.2-54.9%; P > 0.05), before declining significantly at 22 h (32.7%) and 24 h (28.0%). Embryonic arrest was significantly lower in the 16-20 h groups. Blastocysts produced after 16-20 h exhibited higher (P < 0.05) expression of HSPB1, SOD2, GPX3, GDF9, BMP15, STAR, and BCL2, with lower BAX and CASP3 expression, indicating enhanced developmental competence.
Although blastocysts developed following as little as 2 h of gamete co-incubation, a duration of 16 h produced outcomes comparable to the conventional 20-h protocol. Therefore, 16-20 h represents the optimal gamete co-incubation window for maximizing blastocyst yield and developmental competence in buffalo IVF.

PMID:
42489973
Bibliographic data and abstract were imported from PubMed on 23 Jul 2026.

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