Differences in developmental competence and gene expression profiles between buffalo (Bubalus bubalis) preimplantation embryos cultured in three different embryo culture media

Abstract

The objective of this study was to compare effects of in vitro culture systems on embryonic development and expression patterns of developmentally important genes in preimplantation buffalo embryos. After IVM/IVF presumptive zygotes were cultured in one of three systems: undefined TCM-199, mCR2aa medium supplemented with 10 % FBS and defined PVA-myo-inositol-phosphate-EGF medium. No (P [ 0.05) differences at 2-cell, 4-cell and 8-cell to 16-cell stages were observed among the three cultured media used, however, increased (P \ 0.05) blastocyst yield, cell number and hatching rate were found in defined medium compared to undefined media. The expression patterns of genes implicated in embryo metabolism (GLUT-1), anti-apoptosis (BCL-2), imprinting (IGF-2R), DNA methylation (DNMT-3A) and maternal recognition of pregnancy (IFNT) were increased (P \ 0.05) in hatched blastocysts derived from defined medium compared to undefined media. In conclusion, serum-free, defined medium improved developmental competence of in vitro cultured buffalo embryos. Whether these differences in morphological development and gene expression have long-term effects on buffalo calves born after embryo transfer remains unknown. However, it is possible that early adaptations of the preimplantation embryo to its environment persist during fetal and post-natal development.

Cleavage Rate of in vitro Matured Oocytes Fertilized with Conventional Semen in Buffaloes

“…In the present study, average number of blastocyst formed were 3.92±0.46 and blastocyst percentage was 21.57±1.75% as depicted in Table 7 which was in accordance with (Neglia et al, 2003) who observed blastocyst % of 19.9 ± 4.2%. Blastocyst % observed was in accordance with (Ocampo et al, 2016 andSadeesh et al, 2016) as 19.6%, 20.4±2.5% respectively. Blastocyst percentage was higher than present study (Aquino et al, 2013, Ocampo et al, 2015and Shabankareh et al, 2015 as 32.50%, 22.5% and 38.7% respectively and lower than present study (Palta and Chauhan et al, 1998, Boni et al, 1999, Prasad et al, 2013 as 10-15%, 13.5%, 10.65 ± 0.45% respectively in Plate 3.…”

“…Cleavage rates observed (Madan et al, 1994, Mehmood et al, 2007, Suresh et al, 2009and Gonclaves et al, 2013 were higher 54.4%, 75.9%, 52.14 ± 1.15% and 71.8% respectively. Furthermore, cleavage rates observed (Ocampo et al, 2015, Sadeesh et al, 2016 were lower 39.6%, 22.5±4.2% respectively than present study as depicted in Plate 2.…”

Modern achievements of the development of scientific research at obtaining in vitro sheep embryos

“…Так, C. Schwarzer et al (2012) встановили, що 13 різних протоколів культивування ембріонів призводять до чітко виражених клітинних та молекулярних фенотипів у мишачих ембріонів, що дозволяє припустити, що певні компоненти культурального середовища можуть перешкоджати епігенетичній регуляції генів. Дослідження ембріонального геному свідчать, що умови розвитку ембріонів in vitro сьогодні не можуть повністю імітувати in vivo умови експресії мРНК (Sadeesh et al, 2016). Крім того, R. Fernández-Gonzalez et al (2004) спостерігали, що експресія мРНК деяких імпрексуючих генів суттєво порушена в бластоцистах, культивованих у присутності FCS.…”

Retinoic acid improves maturation rate and upregulates the expression of antioxidant-related genes inin vitromatured buffalo (Bubalus bubalis) oocytes

“…The first buffalo calf produced via assisted reproductive technology and in vitro fertilization has been reported in 1991 [3]. Since that date, several studies have been done to examine the influences of different culture conditions on oocyte and embryo development in buffaloes [4], [5]. In addition, several studies were focused on the low cleavage and developmental rates in in vitro produced buffalo embryos compared to cattle.…”