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Abstract: The aim of this study was to determine the effects of IGF-I on the viability and development of porcine presumptive diploid parthenotes developing in vitro. In vitro derived 2-cell parthenotes were cultured to blastocysts with or without exogenous IGF-I in the presence or absence of BSA. The addition of 10 or 100ng/ml IGF-I into NCSU 23 medium did not enhance the development of 2-cell parthenotes to the blastocyst stage. However, IGF-I increased cell numbers (P < 0.001) in the presence of BSA. Similarly, IGF-I alone did not affect apoptosis in the blastocysts but when combined with BSA, it reduced apoptosis (P < 0.05). Real time reverse-transcription polymerase chain reaction revealed that IGF-I significantly decreased the mRNA expression (pro-apoptotic gene Fas or Bak and anti-apoptotic gene Bcl-xL) ratios of both Fas/Bcl-xL (P < 0.01) and Bak/Bcl-xL (P < 0.01) in the presence of 0.4% BSA. These data suggest that IGF-I synergies with BSA to act as a mitogen and apoptotic survival factor in porcine preimplantation parthenotes. [Copyright &y& Elsevier]

The aim of this study was to determine the effects of mouse granulocyte–macrophage colony-stimulating factor (mGM-CSF) on development of porcine parthenotes and nuclear transferred embryos, and on their expression of implantation-related genes. In the presence of bovine serum albumin, mGM-CSF did not increase the percentage of oocytes that developed to the blastocyst stage and at day 7 did not increase cell numbers of embryos. Addition of 2 ng/ml GM-CSF to protein-free culture medium significantly increased the compaction and blastocoel formation of 1- to 2-cell parthenotes developing in vitro. However, total cell numbers were not increased when they were cultured in the presence of GM-CSF. Semi-quantitative reverse transcriptase polymerase chain reaction revealed that mGM-CSF enhances mRNA expression of the leukemia inhibitory factor receptor, but does not influence interleukin-6 or sodium/glucose co-transporter protein gene expression in blastocyst stage parthenotes. These results suggest that mGM-CSF may enhance viability of porcine embryos developing in vitro in a defined culture medium. [Copyright &y& Elsevier]

Abstract: High mobility group box 1 (HMGB1) is considered a component of chromatin and membranes with a role in a variety of biologically important processes. The aim of this study was to determine the effects of HMGB1 on the viability and development of porcine diploid parthenotes cultured in vitro. In vitro derived 4-cell parthenotes were cultured to blastocysts, with or without recombinant HMGB1, in the presence or absence of BSA. The addition of 1, 10, 100 or 1000ng/mL HMGB1 into NCSU 23 medium containing 0.4% BSA did not enhance the development of 4-cell parthenotes to the blastocyst stage and did not change the total number of nuclei in the blastocysts. However, addition of 10 or 100ng/mL HMGB1 into NCSU 23 medium in the absence of BSA increased (P <0.05) both the development rate of parthenotes to the blastocyst stage and total cell numbers. When cultured in NCSU23 medium supplemented with 10 or 100ng/mL HMGB1 and without BSA, apoptosis in parthenotes at the blastocyst stage was decreased (P <0.05). Based on real time RT-PCR, the addition of HMGB1 to the culture medium in the absence of BSA decreased mRNA expression of pro-apoptotic genes Bak (P <0.005) or Caspase3 (Casp3, P <0.01), but not Bcl-xL (Bcl2l2). In conclusion, we inferred that recombinant HMGB1 in the culture medium in the absence of BSA prevented apoptosis of porcine parthenotes and enhanced porcine embryo viability. [Copyright &y& Elsevier]