title: Analysis of imprinted IGF2/H19 gene methylation and expression in normal fertilized and parthenogenetic embryonic stem cells of pigs.
abstract: To determine whether the genomic imprinting can be maintained during the process of embryonic stem (ES) cell derivation from pig blastocysts, mRNA and DNA methylation at the IGF2/H19 imprinting control region in putative ES cells derived from in vitro fertilized (IVF) and parthenogenetic (PG) embryos were investigated. In the present study, one IVF- and three PG ES-like cell lines were established and analyzed for cellular characteristics such as pluripotent marker expression and differentiation capacity. The results showed that these putative ES cells derived from pig blastocysts fulfilled the general "stemness" criteria. The expression of the H19 gene was significantly greater in PG blastocysts than IVF blastocysts, but there were greater amounts of IGF2 in IVF than PG blastocysts. Of these putative ES cell lines, one PG line had less H19 gene expression than a IVF ES cell line while the other two PG lines had much greater expression of the H19 gene than the IVF line. In contrast, the IGF2 gene was upregulated in the same PG cell line relative to the other two PG cell lines and transcript abundance was similar to IVF ES-like cells. Despite the variable amounts of mRNA among the PG cell lines, the IGF2/H19 gene had a differentially methylated region (DMR) 3 was typically un-methylated in all PG cells, and hemi-methylated in the IVF cells. These findings indicated that the mRNA of H19 and IGF2 genes is susceptible to in vitro environments during the process of ES cell derivation from blastocysts but DNA methylation status at this region was well maintained. These altered gene expressions may not be associated with the methylation of the imprinting control region at this locus. Therefore, with their uni-parental genotype, the pluripotent differentiation potentials of PG ES cells could be a valuable tool for understanding genomic imprinting in embryonic development.
authors.affiliation: Department of Agricultural Biotechnology, Animal Biotechnology Major, and Research Institute for Agriculture and Life Science, Seoul National University, Seoul, Republic of Korea; Sooam Biotech Research Foundation, Seoul, Republic of Korea.
authors.forename: Kyung-Jun
authors.initials: KJ
authors.lastname: Uh
authors.affiliation: Sooam Biotech Research Foundation, Seoul, Republic of Korea.
authors.forename: Chi-Hun
authors.initials: CH
authors.lastname: Park
authors.affiliation: Department of Agricultural Biotechnology, Animal Biotechnology Major, and Research Institute for Agriculture and Life Science, Seoul National University, Seoul, Republic of Korea.
authors.forename: Kwang-Hwan
authors.initials: KH
authors.lastname: Choi
authors.affiliation: Department of Agricultural Biotechnology, Animal Biotechnology Major, and Research Institute for Agriculture and Life Science, Seoul National University, Seoul, Republic of Korea.
authors.forename: Jin-Kyu
authors.initials: JK
authors.lastname: Park
authors.affiliation: Sooam Biotech Research Foundation, Seoul, Republic of Korea.
authors.forename: Yeon-Woo
authors.initials: YW
authors.lastname: Jeong
authors.affiliation: Dental Research Institute and CLS21, Seoul National University School of Dentistry, Seoul, Republic of Korea.
authors.forename: Sangho
authors.initials: S
authors.lastname: Roh
authors.affiliation: College of Veterinary Medicine, Chungbuk National University, Cheongju, Republic of Korea.
authors.forename: Sang-Hwan
authors.initials: SH
authors.lastname: Hyun
authors.affiliation: Sooam Biotech Research Foundation, Seoul, Republic of Korea.
authors.forename: Taeyoung
authors.initials: T
authors.lastname: Shin
authors.affiliation: Department of Agricultural Biotechnology, Animal Biotechnology Major, and Research Institute for Agriculture and Life Science, Seoul National University, Seoul, Republic of Korea. Electronic address: leeck@snu.ac.kr.
authors.forename: Chang-Kyu
authors.initials: CK
authors.lastname: Lee
authors.affiliation: Sooam Biotech Research Foundation, Seoul, Republic of Korea. Electronic address: hwangws@sooam.org.
authors.forename: Woo Suk
authors.initials: WS
authors.lastname: Hwang
mesh_terms: D000818:Animals; D001755:Blastocyst; D002460:Cell Line; D019175:DNA Methylation; D053595:Embryonic Stem Cells; D005307:Fertilization in Vitro; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D018392:Genomic Imprinting; D007335:Insulin-Like Growth Factor II / Q000235:genetics / Q000378:metabolism*; D059785:Karyotype; D010312:Parthenogenesis; D062085:RNA, Long Noncoding / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013552:Swine / Q000502:physiology*
chemical_list: C094335:H19 long non-coding RNA; D062085:RNA, Long Noncoding; D012333:RNA, Messenger; D007335:Insulin-Like Growth Factor II
keywords: DNA methylation; Embryonic stem cell; Imprinted gene; Parthenogenesis