title: Porcine granulocyte-macrophage colony-stimulating factor improves the in vitro development of cloned porcine embryos.
abstract: We examined the effects of porcine granulocyte-macrophage colony-stimulating factor (pGM-CSF) on the in vitro development of porcine embryos produced by somatic cell nuclear transfer (SCNT) for the first time. We evaluated the effects of pGM-CSF on SCNT-derived blastocyst formation and investigated gene expression. A total of 522 cloned embryos in 6 replicates were treated with 10 ng/ml pGM-CSF during in vitro culture (IVC). This treatment significantly (P<0.05) increased blastocyst formation and total cell number in blastocysts compared with the control (12.3% and 41.4 vs. 9.0% and 34.7, respectively). However, there was no effect on cleavage rate. The numbers of cells in the inner cell mass and trophectoderm were significantly higher in the pGM-CSF treatment group (6.0 and 43.0, respectively) compared with the control (4.4 and 31.9, respectively). Treatment with 10 ng/ml pGM-CSF significantly increased POU5F1 and Cdx2 mRNA expression in blastocysts. In addition, Bcl-2, Dnmt1 and proliferating cell nuclear antigen (PCNA) mRNA expression were upregulated in blastocysts in the pGM-CSF supplemented group compared with the control. These results suggest that pGM-CSF improves the quality and developmental viability of porcine SCNT embryos by regulating transcription factor expression.
authors.affiliation: Laboratory of Veterinary Embryology and Biotechnology, College of Veterinary Medicine, Chungbuk National University, Cheongju 361-763, Republic of Korea.
authors.forename: Seong-Sung
authors.initials: SS
authors.lastname: Kwak
authors.forename: Seung-A
authors.initials: SA
authors.lastname: Cheong
authors.forename: Yubyeol
authors.initials: Y
authors.lastname: Jeon
authors.forename: Sang-Hwan
authors.initials: SH
authors.lastname: Hyun
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000187:drug effects*; D000071616:CDX2 Transcription Factor; D016022:Case-Control Studies; D017931:DNA Primers / Q000235:genetics; D046149:Embryo Culture Techniques / Q000662:veterinary; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology*; D018398:Homeodomain Proteins / Q000378:metabolism; D053652:Nuclear Transfer Techniques / Q000662:veterinary; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary; D034421:Sus scrofa / Q000196:embryology*; D015534:Trans-Activators / Q000378:metabolism
chemical_list: D000071616:CDX2 Transcription Factor; D017931:DNA Primers; D018398:Homeodomain Proteins; D050814:Octamer Transcription Factor-3; D018809:Proliferating Cell Nuclear Antigen; D019253:Proto-Oncogene Proteins c-bcl-2; D015534:Trans-Activators; D016178:Granulocyte-Macrophage Colony-Stimulating Factor