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FACS selection of valuable mutant mouse round spermatids and strain rescue via round spermatid injection

Published online by Cambridge University Press:  20 December 2013

Lian Zhu ,
Wei Zhou ,
Peng-Cheng Kong ,
Mei-Shan Wang ,
Yan Zhu ,
Li-Xin Feng ,
Xue-Jin Chen  and
Man-Xi Jiang
Lian Zhu
Affiliation:
Department of Laboratory Animal Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
Wei Zhou
Affiliation:
Department of Developmental Biology, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
Peng-Cheng Kong
Affiliation:
Department of Laboratory Animal Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
Mei-Shan Wang
Affiliation:
Department of Laboratory Animal Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China. College of Wildlife Resource, Northeast Forestry University, Harbin 150040, China.
Yan Zhu
Affiliation:
Key Laboratory of Contraceptive Drugs and Devices of National Population and Family Planning Committee, Shanghai Institute of Planned Parenthood Research, Shanghai 200032, China.
Li-Xin Feng
Affiliation:
Department of Developmental Biology, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
Xue-Jin Chen*
Affiliation:
Department of Laboratory Animal Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China. Department of Laboratory Animal Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
Man-Xi Jiang*
Affiliation:
Department of Laboratory Animal Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China. Department of Laboratory Animal Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
*
All correspondence to: Man-Xi Jiang or Xue-Jin Chen. Department of Laboratory Animal Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China. Tel: +86 21 63846590, ext. 776539. e-mail: manxijiang@yahoo.com or chenxuejin@shsmu.edu.cn
All correspondence to: Man-Xi Jiang or Xue-Jin Chen. Department of Laboratory Animal Sciences, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China. Tel: +86 21 63846590, ext. 776539. e-mail: manxijiang@yahoo.com or chenxuejin@shsmu.edu.cn
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Summary

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.

Keywords

MouseFACSROSIRound spermatidsStrain rescue
Type
Research Article
Information
Zygote , Volume 23 , Issue 3 , June 2015 , pp. 336 - 341
Copyright
Copyright © Cambridge University Press 2013 

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