Development of buffalo embryo derived from ICSI: effects of various somatic-cell co-culture

ContextSomatic-cell co-culture of intracytoplasmic sperm injection (ICSI) buffalo embryos has not been reported earlier.AimThis study aimed to determine the effects of buffalo oviductal epithelial-cell, granulosa-cell, and cumulus-cell co-culture on in vitro culture of early embryo development as ICSI and post-activation.MethodsSelected oocyte-cumulus complexes were cultured for 19-20 h in 50-mu L drop of tissue culture medium (TCM199 + 10% buffalo follicular fluid, hCG 50 IU/mL, 0.02% arbitrary units (AU)/mL follicle-stimulating hormone and 1 mu g/mL estradiol-17 beta E2). Oocytes reaching Metaphase II were subjected to ICSI with immobilised spermatozoa. All ICSI oocytes were activated with calcium ionophore for 5 min, followed by cycloheximide for 5 h. The embryos at 6-8-cell stages were co-cultured.Key resultsThe morula, blastocyst, and hatched blastocyst rates when co-cultured with oviductal epithelial cells were 68.18%, 48.18%, and 30.00% respectively. The morula, blastocyst, and hatched blastocyst rates when co-cultured with cumulus cells were 51.49%, 34.33%, and 16.42% respectively. The morula, blastocyst, and hatched blastocyst rates when co-cultured with granulosa cells were 52.14%, 32.48%, and 13.68% respectively.ConclusionsIn vitro maturation buffalo oocytes can be fertilised in vitro with ICSI and co-cultured with different types of cells. Oviductal epithelial cell co-culture was shown to be superior in supporting in vitro embryo development in this study.ImplicationsThe oviductal epithelial cells are easy to prepare and may be used for co-culture to increase the efficiency of in vitro production of buffalo embryos. The buffalo is an important livestock resource in the agricultural system. The low efficiency of buffalo reproduction needs to be overcome. Sperm-injection technique and an increased production efficiency of in vitro embryos in somatic cell co-culture are among the technologies that can assist, enhance and increase the population of buffaloes.