title: FACS selection of valuable mutant mouse round spermatids and strain rescue via round spermatid injection.
abstract: Round spermatid injection (ROSI) into mammalian oocytes can result in the development of viable embryos and offspring. One current limitation to this technique is the identification of suitable round spermatids. In the current paper, round spermatids were selected from testicular cells with phase contrast microscopy (PCM) and fluorescence-activated cell sorting (FACS), and ROSI was performed in two strains of mice. The rates of fertilization, embryonic development and offspring achieved were the same in all strains. Significantly, round spermatids selected by PCM and FACS were effectively used to rescue the infertile Pten-null mouse. The current results indicate that FACS selection of round spermatids can not only provide high-purity and viable round spermatids for use in ROSI, but also has no harmful effects on the developmental capacity of subsequently fertilized embryos. It was concluded that round spermatids selected by FACS are useful for mouse strain rederivation and rescue of infertile males; ROSI should be considered as a powerful addition to the armamentarium of assisted reproduction techniques applicable in the mouse.
authors.affiliation: Department of Laboratory Animal Sciences,Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China.
authors.forename: Lian
authors.initials: L
authors.lastname: Zhu
authors.affiliation: Department of Developmental Biology,Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhou
authors.affiliation: Department of Laboratory Animal Sciences,Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China.
authors.forename: Peng-Cheng
authors.initials: PC
authors.lastname: Kong
authors.affiliation: Department of Laboratory Animal Sciences,Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China.
authors.forename: Mei-Shan
authors.initials: MS
authors.lastname: Wang
authors.affiliation: Key Laboratory of Contraceptive Drugs and Devices of National Population and Family Planning Committee,Shanghai Institute of Planned Parenthood Research,Shanghai 200032,China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhu
authors.affiliation: Department of Developmental Biology,Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China.
authors.forename: Li-Xin
authors.initials: LX
authors.lastname: Feng
authors.affiliation: Department of Laboratory Animal Sciences,Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China.
authors.forename: Xue-Jin
authors.initials: XJ
authors.lastname: Chen
authors.affiliation: Department of Laboratory Animal Sciences,Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China.
authors.forename: Man-Xi
authors.initials: MX
authors.lastname: Jiang
mesh_terms: D000818:Animals; D004624:Embryo Transfer; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D008297:Male; D008813:Mice, Inbred ICR; D008817:Mice, Mutant Strains; D008822:Mice, Transgenic; D008858:Microscopy, Phase-Contrast; D051059:PTEN Phosphohydrolase / Q000235:genetics; D011247:Pregnancy; D018873:Pregnancy Rate; D020554:Sperm Injections, Intracytoplasmic / Q000379:methods*; D013087:Spermatids / Q000166:cytology* / Q000502:physiology; D013737:Testis / Q000166:cytology
chemical_list: D051059:PTEN Phosphohydrolase; C487757:Pten protein, mouse
keywords: Strain rescue