title: Supplementation With Cell-Penetrating Peptide-Conjugated Estrogen-Related Receptor β Improves the Formation of the Inner Cell Mass and the Development of Vitrified/Warmed Mouse Embryos.
abstract: Estrogen-related receptor β (ESRRB), which is a member of the nuclear orphan receptor family, regulates the messenger RNA (mRNA) expression levels of the transcription factors, Oct4 and Nanog, in early embryos and germ cells, thereby maintaining the undifferentiated state and pluripotency of the relevant cells. The present study was designed to determine whether the upregulation of pluripotency-related genes by direct delivery of ESRRB protein may affect on the commitment into inner cell mass (ICM) or the development of vitrified/warmed mouse embryos. Recombinant cell-penetrating peptide (CPP) ESRRB protein was synthesized and then added into a culture medium for cryopreserved mouse embryos. Vitrified/warmed 8-cell embryos were cultured in KSOM with/without 2 μg/mL CPP-ESRRB for 48 hours and then analyzed or transferred to the uteri of foster mothers. The mRNA expression of Oct4 and Nanog was higher in CPP-ESRRB-treated blastocysts compared to the untreated controls. No difference was observed in embryonic development, but ICM:trophectoderm ratio was increased in the CPP-ESRRB-treated group compared to the untreated group, and after embryo transfer, a higher implantation rate was obtained in the CPP-ESRRB-treated group compared to the untreated group. This study shows for the first time that recombinant CPP-ESRRB can be easily integrated into vitrified/warmed mouse embryos and that it increases Oct4 expression (via a pluripotency-related gene pathway), ICM formation, and the further embryonic and full-term development of vitrified/warmed mouse embryos. This CPP-conjugated protein delivery system could therefore be a useful tool for improving assisted reproductive technology.
authors.affiliation: Department of Biomedical Science, College of Life Science, CHA University, Seongnam-si, Korea.
authors.forename: Ning Jie
authors.initials: NJ
authors.lastname: Yang
authors.affiliation: Department of Biomedical Science, College of Life Science, CHA University, Seongnam-si, Korea.
authors.forename: Dong-Won
authors.initials: DW
authors.lastname: Seol
authors.affiliation: Department of Biomedical Science, College of Life Science, CHA University, Seongnam-si, Korea.
authors.forename: Junghyun
authors.initials: J
authors.lastname: Jo
authors.affiliation: Department of Biomedical Science, College of Life Science, CHA University, Seongnam-si, Korea.
authors.forename: Hyun Mee
authors.initials: HM
authors.lastname: Jang
authors.affiliation: Department of Biomedical Science, College of Life Science, CHA University, Seongnam-si, Korea Fertility Center, CHA Gangnam Medical Center, College of Medicine, CHA University, Seoul, Korea.
authors.forename: Sook-Young
authors.initials: SY
authors.lastname: Yoon
authors.affiliation: Fertility Center, CHA Gangnam Medical Center, College of Medicine, CHA University, Seoul, Korea.
authors.forename: Woo Sik
authors.initials: WS
authors.lastname: Lee
authors.affiliation: Department of Biomedical Science, College of Life Science, CHA University, Seongnam-si, Korea Fertility Center, CHA Gangnam Medical Center, College of Medicine, CHA University, Seoul, Korea drleedr@cha.ac.kr wooslee@cha.ac.kr.
authors.forename: Dong Ryul
authors.initials: DR
authors.lastname: Lee
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000187:drug effects* / Q000378:metabolism*; D002452:Cell Count; D057846:Cell-Penetrating Peptides / Q000008:administration & dosage*; D003470:Culture Media; D010064:Embryo Implantation / Q000187:drug effects; D004624:Embryo Transfer; D047108:Embryonic Development; D004966:Estrogens, Conjugated (USP) / Q000008:administration & dosage*; D005260:Female; D018507:Gene Expression Regulation, Developmental; D008297:Male; D051379:Mice; D000071317:Nanog Homeobox Protein / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D011960:Receptors, Estrogen / Q000008:administration & dosage*; D058989:Vitrification
chemical_list: D057846:Cell-Penetrating Peptides; D003470:Culture Media; C098925:Esrrb protein, mouse; D004966:Estrogens, Conjugated (USP); D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D012333:RNA, Messenger; D011960:Receptors, Estrogen
keywords: 
  Oct4
  ; CPP-ESRRB; blastocyst formation; embryo development; inner cell mass; mouse; vitrification