title: Birth of healthy offspring following ICSI in in vitro-matured common marmoset (Callithrix jacchus) oocytes.
abstract: Intracytoplasmic sperm injection (ICSI), an important method used to treat male subfertility, is applied in the transgenic technology of sperm-mediated gene transfer. However, no study has described successful generation of offspring using ICSI in the common marmoset, a small non-human primate used as a model for biomedical translational research. In this study, we investigated blastocyst development and the subsequent live offspring stages of marmoset oocytes matured in vitro and fertilized by ICSI. To investigate the optimal timing of performing ICSI, corrected immature oocytes were matured in vitro and ICSI was performed at various time points (1-2 h, 2-4 h, 4-6 h, 6-8 h, and 8-10 h after extrusion of the first polar body (PB)). Matured oocytes were then divided randomly into two groups: one was used for in vitro fertilization (IVF) and the other for ICSI. To investigate in vivo development of embryos followed by ICSI, 6-cell- to 8-cell-stage embryos and blastocysts were nonsurgically transferred into recipient marmosets. Although no significant differences were observed in the fertilization rate of blastocysts among ICSI timing after the first PB extrusion, the blastocyst rate at 1-2 h was lowest among groups at 2-4 h, 4-6 h, 6-8 h, and 8-10 h. Comparing ICSI to IVF, the fertilization rates obtained in ICSI were higher than in IVF (p>0.05). No significant difference was noted in the cleaved blastocyst rate between ICSI and IVF. Following the transfer of 37 ICSI blastocysts, 4 of 20 recipients became pregnant, while with the transfer of 21 6-cell- to 8-cell-stage ICSI embryos, 3 of 8 recipients became pregnant. Four healthy offspring were produced and grew normally. These are the first marmoset offspring produced by ICSI, making it an effective fertilization method for marmosets.
authors.affiliation: Department of Applied Developmental Biology, Central Institute for Experimental Animals, Kawasaki-ku, Kawasaki, Kanagawa, Japan; Department of Biochemistry, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan.
authors.forename: Tsukasa
authors.initials: T
authors.lastname: Takahashi
authors.affiliation: Department of Oncology, Juntendo University Nerima Hospital, Nerima-ku, Tokyo, Japan.
authors.forename: Kisaburo
authors.initials: K
authors.lastname: Hanazawa
authors.affiliation: Marmoset Research Department, Central Institute for Experimental Animals, Kawasaki-ku, Kawasaki, Kanagawa, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Inoue
authors.affiliation: Department of Applied Developmental Biology, Central Institute for Experimental Animals, Kawasaki-ku, Kawasaki, Kanagawa, Japan.
authors.forename: Kenya
authors.initials: K
authors.lastname: Sato
authors.affiliation: Department of Applied Developmental Biology, Central Institute for Experimental Animals, Kawasaki-ku, Kawasaki, Kanagawa, Japan.
authors.forename: Ayako
authors.initials: A
authors.lastname: Sedohara
authors.affiliation: Department of Applied Developmental Biology, Central Institute for Experimental Animals, Kawasaki-ku, Kawasaki, Kanagawa, Japan.
authors.forename: Junko
authors.initials: J
authors.lastname: Okahara
authors.affiliation: Department of Biomedical Research, Central Institute for Experimental Animals, Kawasaki-ku, Kawasaki, Kanagawa, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Suemizu
authors.affiliation: Department of Biomedical Research, Central Institute for Experimental Animals, Kawasaki-ku, Kawasaki, Kanagawa, Japan.
authors.forename: Chie
authors.initials: C
authors.lastname: Yagihashi
authors.affiliation: Department of Biomedical Research, Central Institute for Experimental Animals, Kawasaki-ku, Kawasaki, Kanagawa, Japan.
authors.forename: Masafumi
authors.initials: M
authors.lastname: Yamamoto
authors.affiliation: Department of Applied Developmental Biology, Central Institute for Experimental Animals, Kawasaki-ku, Kawasaki, Kanagawa, Japan.
authors.forename: Tomoo
authors.initials: T
authors.lastname: Eto
authors.affiliation: Altair Corporation, Kohoku-ku, Yokohama-shi, Kanagawa, Japan.
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Konno
authors.affiliation: Department of Physiology, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan.
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Okano
authors.affiliation: Department of Biochemistry, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Suematsu
authors.affiliation: Department of Applied Developmental Biology, Central Institute for Experimental Animals, Kawasaki-ku, Kawasaki, Kanagawa, Japan; Keio Advanced Research Center, Keio University, Shinjuku-ku, Tokyo, Japan.
authors.forename: Erika
authors.initials: E
authors.lastname: Sasaki
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001755:Blastocyst / Q000166:cytology; D002144:Callithrix / Q000196:embryology*; D002454:Cell Differentiation; D004624:Embryo Transfer; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism; D047108:Embryonic Development; D005260:Female; D005819:Genetic Markers; D006801:Humans; D008297:Male; D018895:Microsatellite Repeats / Q000235:genetics; D009865:Oocytes / Q000378:metabolism*; D059705:Polar Bodies / Q000166:cytology; D011247:Pregnancy; D020554:Sperm Injections, Intracytoplasmic / Q000379:methods*
chemical_list: D005819:Genetic Markers
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