Gene Expression and Developmental Competence of Bovine Embryos Produced In Vitro with Different Serum Concentrations

Abstract

The objective of this study was to determine the developmental rates and relative abundance of Hsp 70.1 and Glut-1 transcripts in in vivo- and in vitro-produced (IVP) bovine embryos in media supplemented with bovine serum albumin (BSA) or different oestrous cow serum concentrations. In experiment 1, in vitro maturation and culture media were supplemented with 0.4% BSA or 1, 5, 10 or 20% of oestrous cow serum (ECS). The analysis of the expression of Hsp 70.1 and Glut-1 was carried out in individual days 7 and 8 embryos by a semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) assay. In experiment 2, in vivo-produced morulae were collected on day 7 of the oestrous cycle and employed for the comparison of the relative abundances of Hsp 70.1 and Glut-1 transcripts with IVP morulae produced using two protein sources (10% ECS or 0.4% BSA). No differences were observed in cleavage rate among groups, but blastocyst formation (27%) and hatching rates (78%) were significantly higher in IVP embryos produced with 20% ECS than the other groups (p<0.05). No significant differences were observed in the relative abundances of Hsp 70.1 and Glut-1 mRNA in days 7 and 8 blastocysts expanded blastocysts between groups. The abundances of mRNA for those genes were similar between IVP and in vivo-produced morulae. In spite of the alterations observed in embryonic development, the presence of serum at distinct concentrations did not appear to alter the relative abundance profiles of Hsp 70.1 and Glut-1 compared with controls or the BSA supplementation to the IVP media.

Expression and Characterization of Constitutive Heat Shock Protein 70.1 (HSPA‐1A) Gene in In Vitro Produced and In Vivo‐Derived Buffalo (Bubalus bubalis) Embryos

“…The HSPA‐1A confers thermotolerance against variety of stressors like chemical, environmental and nutritional, and it can be used as stress indicator in bovine embryos (Wrenzycki et al. 2001; Oliveira et al. 2006).…”

Efficiency of SCNT Buffalo (Bubalus bubalis) Embryos in Different Culture Medium and Analysis of mRNA Expression of Insulin-like Growth Factors During Embryogenesis

“…They are able to develop in media ranging in composition from simple balanced salt solutions to complex systems supplemented with serum and somatic cells (Lonergan et al. 1999; de Oliveira et al. 2006).…”

A microfluidic in vitro cultivation system for mechanical stimulation of bovine embryos

“…To improve the developmental efficiency and quality of embryos, many studies have addressed the chemical modification of medium [5] and its components, as well as environmental changes in physiological conditions, such as temperature [6], pH, shear stress [7], and osmolality [8]. Although these efforts have revealed many factors that are critical to developing embryos, the development rate of embryos, especially that of large animals such as porcine and bovine species, still remains low [9]. This low efficiency is believed to be due to the different environments of in vitro and in vivo conditions.…”