title: Generation of viable embryos and embryonic stem cell-like cells from cultured primary follicles in mice.
abstract: Primary follicles retrieved from B6CBAF1 prepubertal mice were cultured in a stepwise manner in an alpha-minimum essential medium-based medium to generate viable embryos and embryonic stem cell (ESC)-like cells. A significant increase in follicle growth and oocyte maturation accompanied by increased secretion of 17beta-estradiol and progesterone was achieved by exposing primary follicles to 100 or 200 mIU of follicle-stimulating hormone (FSH) during culture. More oocytes developed into blastocysts following in vitro fertilization (IVF) or parthenogenetic activation after culture with 200 mIU of FSH during the entire culture period than with 100 mIU. Eleven ESC-like cell lines, consisting of four heterozygotic and seven homozygotic phenotypes, were established from 25 trials of primary follicle culture combined with IVF or parthenogenetic activation. In conclusion, primary follicles can potentially yield developmentally competent oocytes, which produce viable embryos and ESC-like cell lines following in vitro manipulation. We suggest a method to utilize immature follicles, which are most abundant in ovaries, to improve reproductive efficiency and for use in regenerative medicine.
authors.affiliation: Laboratory of Stem Cell and Bioevaluation, World Class University Biomodulation Program, Seoul National University, Seoul, Republic of Korea.
authors.forename: Jun Hee
authors.initials: JH
authors.lastname: Choi
authors.forename: Gil Ah
authors.initials: GA
authors.lastname: Kim
authors.forename: Jong Heum
authors.initials: JH
authors.lastname: Park
authors.forename: Gwon Hwa
authors.initials: GH
authors.lastname: Song
authors.forename: Jun Won
authors.initials: JW
authors.lastname: Park
authors.forename: Dae Yong
authors.initials: DY
authors.lastname: Kim
authors.forename: Jeong Mook
authors.initials: JM
authors.lastname: Lim
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D001755:Blastocyst / Q000166:cytology* / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival; D003433:Crosses, Genetic; D004477:Ectogenesis; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D004958:Estradiol / Q000378:metabolism; D005260:Female; D005307:Fertilization in Vitro; D005640:Follicle Stimulating Hormone / Q000378:metabolism; D059471:In Vitro Oocyte Maturation Techniques; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D009866:Oogenesis; D006080:Ovarian Follicle / Q000166:cytology* / Q000378:metabolism; D010312:Parthenogenesis; D011374:Progesterone / Q000378:metabolism; D046509:Tissue Culture Techniques; D014022:Tissue Survival
chemical_list: D015415:Biomarkers; D011374:Progesterone; D004958:Estradiol; D005640:Follicle Stimulating Hormone