title: Effect of primary culture medium type for culture of canine fibroblasts on production of cloned dogs.
abstract: Fibroblasts are common source of donor cells for SCNT. It is suggested that donor cells' microenvironment, including the primary culture, affects development of reconstructed embryos. To prove this, canine embryos were cloned with fibroblasts that were cultured in two different primary media (RCMEp vs. Dulbecco's modified Eagle's medium [DMEM]) and in vivo developments were compared with relative amount of stemness, reprogramming, apoptosis gene transcripts, and telomerase activity. Donor cells cultured in RCMEp contained a significantly higher amount of SOX2, NANOG, DPPA2, REXO1, HDAC, DNMT1, MECP2 and telomerase activity than those cultured in DMEM (P < 0.05). In vivo developmental potential of cloned embryos with donor cells cultured in RCMEp had a higher birth rate than that of embryos derived from DMEM (P < 0.05). The culture medium can induce changes in gene expression of donor cells and telomerase activity, and these alterations can also affect in vivo developmental competence of the cloned embryos.
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Seoul, Korea.
authors.forename: Geon A
authors.initials: GA
authors.lastname: Kim
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Seoul, Korea.
authors.forename: Hyun Ju
authors.initials: HJ
authors.lastname: Oh
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Seoul, Korea.
authors.forename: Min Jung
authors.initials: MJ
authors.lastname: Kim
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Seoul, Korea.
authors.forename: Young Kwang
authors.initials: YK
authors.lastname: Jo
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Seoul, Korea.
authors.forename: Jin
authors.initials: J
authors.lastname: Choi
authors.affiliation: College of Veterinary Medicine, Chungbuk National University, Cheongju, Korea.
authors.forename: Jin Wook
authors.initials: JW
authors.lastname: Kim
authors.affiliation: College of Natural and Mathematical Sciences, University of Maryland Baltimore County, Baltimore, Maryland, USA.
authors.forename: Tae Hee
authors.initials: TH
authors.lastname: Lee
authors.affiliation: Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Seoul, Korea. Electronic address: bclee@snu.ac.kr.
authors.forename: Byeong Chun
authors.initials: BC
authors.lastname: Lee
mesh_terms: D000818:Animals; D019976:Cloning, Organism / Q000379:methods / Q000662:veterinary*; D003470:Culture Media; D004285:Dogs / Q000196:embryology* / Q000235:genetics*; D005347:Fibroblasts / Q000187:drug effects* / Q000502:physiology; D018507:Gene Expression Regulation, Developmental / Q000502:physiology; D053652:Nuclear Transfer Techniques / Q000662:veterinary*; D059467:Transcriptome
chemical_list: D003470:Culture Media
keywords: Culture medium; Dog cloning; Primary culture; SCNT