Viability and Growth of Buffalo Preantral Follicles and their Corresponding Oocytes In Vitro: Effect of Growth Factors and β Mercaptoethanol

Abstract

The present study was undertaken to isolate buffalo preantral follicles (PFs), to test the viability and sizes of buffalo PFs and to examine the effect of various growth factors (insulin-like growth factor, fibroblast growth factor) and an antioxidant (beta mercaptoethanol) on the in vitro growth, survival and antrum formation rates of buffalo PFs and growth rates of oocytes in cultured PFs. Preantral follicles from slaughtered buffalo ovaries were recovered by a combined mechanical and enzymatic method. The recovery rates of >40-100, 101-200, 201-300, 301-400 and 401-500 microm PFs were 5.1, 3.2, 3.1, 6.3 and 5.1 per ovary, respectively. The corresponding viability rates were 76.1%, 78.1%, 85.2%, 92.5% and 92.6%, respectively. There was a positive correlation (r = 0.73) between oocyte size and the follicular size. However, there was no significant correlation between the size of oocyte and its viability at the time of its retrieval from ovary. Insulin-like growth factor and fibroblast growth factor improved the survival of buffalo PFs and regulated their growth in culture. The growth factors and beta mercaptoethanol in association synergically improved the growth and survival of buffalo PFs.

Short-Term Culture of Ovarian Cortical Strips From Capuchin Monkeys (Sapajus apella): A Morphological, Viability, and Molecular Study of Preantral Follicular Development In Vitro

“…Ovarian fragments were placed in 500 mL of culture medium in 24-well plates for 24 hours. The basic culture medium consisted of TCM-199 (pH 7.2-7.4) supplemented with 10% fetal calf serum (FCS), 100 ng/mL epidermal growth factor (control for cultured follicles), 1.0 mg/mL insulin, 5.5 mg/mL transferrin, and 5.0 ng/mL selenium (ITS), either or not added with 10 mmol/L BME, 15 100 ng/mL BMP4, 16 or 25 IU PMSG 17 alone or in combination. After IVC, all fragments were divided into 2 equal parts and submitted to follicular isolation or quantitative reverse transcriptase polymerase chain reaction (qRT-PCR).…”

“…To improve preantral follicular survival and development during IVC, medium has been supplemented with reducing agents such as β-mercaptoethanol (BME), 15 growth factors such as BMP4, 16 and a glycoprotein hormone such as pregnant mare serum gonadotrophin (PMSG), which has both follicle-stimulating harmone (FSH-) and lutenizing harmone (LH)-like activities. 17 Except for the studies such as those from Nilsson and Skinner, 16 Abir et al, 12 and Silva et al, 18 assessment of follicular development is mainly or exclusively based on morphological and viability parameters.…”

“…17 Except for the studies such as those from Nilsson and Skinner, 16 Abir et al, 12 and Silva et al, 18 assessment of follicular development is mainly or exclusively based on morphological and viability parameters. 15 ,17 Viable and morphologically normal follicles are, however, not unconditionally able to further develop into more advanced stages, since they might be functionally unable to do so. Therefore, the aim of the present study was to assess the viability, the development, and the molecular profile of in vitro cultured capuchin preantral follicles.…”

Isolation and Culture of Preantral Follicles for Retrieving Oocytes for the Embryo Production: Present Status in Domestic Animals

“…2006). In a recent study, the growth factors (IGF and FGF) and β‐mercaptoethanol in association synergically improved the growth and survival of large buffalo PFs (Gupta and Nandi 2010).…”

Effect of animal-sourced bioactive peptides on the in vitro development of mouse preantral follicles

“…Therefore, the addition of antioxidants to follicle cultures as ROS scavengers could prevent or decrease harmful damages, resulting in increased qualitiy of germ cells [ 14 – 16 ]. Although different antioxidants, such as rutin [ 17 ], β-mercaptoethanol [ 18 ] and α lipoic acid [ 19 ], have been routinely applied to the culture system of developing follicles, the effect of bioactive peptides (BAPT), natural antioxidants derived from various food or animals sources [ 20 ] on the development of mammalian preantral follicles has not yet been investigated.…”