In vitro production of a caprine embryo from a preantral follicle cultured in media supplemented with growth hormone

Abstract

The objective was to evaluate the effects of growth hormone (GH) on the survival, growth, maturation, and fertilization of oocytes derived from caprine preantral ovarian follicles cultured in vitro. Preantral follicles were isolated from the cortex of caprine ovaries and individually cultured for 18 d in the absence (control) or presence of bovine GH at concentrations of 10 or 50 ng/mL (GH10 and GH50, respectively). Follicle development was evaluated on the basis of survival, antral cavity formation, diameter increase, and the presence of healthy cumulus-oocyte complexes and mature oocytes. After culture, oocytes were subjected to in vitro maturation (IVM) and in vitro fertilization (IVF). The rate of antrum formation after Day 6 of culture was higher in both GH10 and GH50 than in the control (81.0, 92.7, and 47.6%, respectively, P < 0.05). Percentages of grown oocytes that were acceptable for IVM were also higher (P < 0.05) in GH-treated groups than in the control (54.8, 48.8, and 11.9% for GH10, GH50, and Control). A higher percentage of oocytes in the GH50 treatment underwent meiotic resumption (50.0%), produced mature oocytes, and enabled production of an embryo after IVF than in the control group (0.0%; P < 0.05). In conclusion, GH promoted in vitro growth and maturation of goat preantral follicle oocytes and enabled production of an embryo. Furthermore, this study was apparently the first to produce a caprine embryo by in vitro fertilization of oocytes derived from preantral follicles grown in vitro.

Effect of Different Combinations of the Growth Factors and Hormones on In vitro Maturation of Goat Preantral Follicles

“…Zhou and Zhang (2005) who reported that, in presence of FSH, IGF-I in combination with EGF produced a higher survival rate of adult caprine preantral follicles than any other growth factor did individually. Findings of present study are in accordance with findings of Magalhaes et al, (2011b) in high rate of follicular survival and antrum formation when media supplemented with Growth hormone (GH). Furthermore, the influence of the thyroxin on PFs appears to depend on the physiological status of the PFs at the time of collection (Gupta et al, 2007).…”

“…In IVM media oocytes obtained from T 3 treatment (T 4 +FSH+GH+EGF) had maximum percentage of M-II stage oocytes (38.6%) in contrast to the findings of Arunakumari et al, (2010) in sheep, the difference might be due to variation in culture media or species difference. However, these findings were in accordance with the Magalhaes et al, (2011b) on production of matured oocytes by supplementation of Growth hormone at a concentration of 10 ng/ml and 50 ng/ml. The present findings were in accordance with the results of Saraiva et al, (2012) who studied the effect of both FSH and LH on oocyte maturation.…”

“…However among farm animals, success has been achieved up to embryo development using oocytes from cultured PFs of adult ovaries in pig (Wu et al, 2001a and b), buffalo (Gupta et al, 2008), sheep (Arunakumari et al, 2010) and goat (Magalhaes et al, 2011a) but none of these studies progressed to develop blastocyst except pig. Addition of GH (50ng/ml) resulted in growth of oocytes form goat preantral follicles that were acceptable for IVM and IVF Magalhaes et al, (2011b). Addition of VEGF-A 165 to the culture medium improved the development of goat PFs cultured in vitro, allowing the production of mature oocytes .…”

Vascular endothelial growth factor-A165 (VEGF-A165) stimulates the in vitro development and oocyte competence of goat preantral follicles

“…As the follicles grow, they need a higher nutrient intake and become dependent on gonadotropins. Some in vitro studies of preantral secondary follicles suggest that these follicles are able to generate competent oocytes that are able to undergo subsequent embryo development in vitro (Wu et al 2001;Gupta et al 2008;Magalhães et al 2010;Arunakumari et al 2010). However, the mechanisms that control the regulation of the in vitro growth of preantral follicules are not completely understood, particularly those that control the development of large secondary follicles.…”

Generation of Embryonic Stem Cell Lines from Immature Rabbit Ovarian Follicles

“…In other species it is possible for embryos to develop to morula or blastocyst stages following in vitro culture of immature follicles [7][8][9][10][11][12]; however, these embryos do not progress to later developmental stages.…”