Technology Report

Generation of progeny from embryonic stem cells by microinsemination of male germ cells from chimeric mice

  1. Eiji Mizutani1,2,*,
  2. Hiroshi Ohta1,
  3. Satoshi Kishigami1,
  4. Nguyen Van Thuan1,
  5. Takafusa Hikichi1,
  6. Sayaka Wakayama1,3,
  7. Eimei Sato2,
  8. Teruhiko Wakayama1

Article first published online: 16 AUG 2005

DOI: 10.1002/gene.20153

Issue

genesis

genesis

Volume 43, Issue 1, pages 34–42, September 2005

Additional Information

How to Cite

Mizutani, E., Ohta, H., Kishigami, S., Van Thuan, N., Hikichi, T., Wakayama, S., Sato, E. and Wakayama, T. (2005), Generation of progeny from embryonic stem cells by microinsemination of male germ cells from chimeric mice. Genesis, 43: 34–42. doi: 10.1002/gene.20153

Author Information

  1. 1

    Laboratory for Genomic Reprogramming, Center for Developmental Biology, Riken Kobe, Japan

  2. 2

    Laboratory of Animal Reproduction, Graduate School of Agricultural Science, Tohoku University, Sendai, Japan

  3. 3

    Department of Life Science, Graduate School of Science and Technology, Kobe University, Kobe, Japan

*Laboratory for Genomic Reprogramming, Center for Developmental Biology, Riken Kobe, Japan

Publication History

  1. Issue published online: 16 AUG 2005
  2. Article first published online: 16 AUG 2005
  3. Manuscript Accepted: 23 JUN 2005
  4. Manuscript Received: 1 APR 2005

Funded by

  • Ministry of Education, Science, Sports, Culture and Technology of Japan. Grant Numbers: Grants-in-Aid for Creative Scientific Research 13GS0008, Scientific Research in Priority Areas 15080211, Young Scientists A 15681014, Project for the Realization of Regenerative Medicine (the research field for technical development of stem cell manipulation)

SEARCH

SEARCH BY CITATION

ARTICLE TOOLS

Get PDF (755K)

Keywords:

  • chimera;
  • GFP;
  • embryonic stem cell;
  • testis;
  • germ cell;
  • germ-line transmission;
  • microinsemination;
  • transgenic mouse

Abstract

Mice chimeric for embryonic stem (ES) cells have not always successfully produced ES-derived offspring. Here we show that the male gametes from ES cells could be selected in male chimeric mice testes by labeling donor ES cells or host blastocytes with GFP. Male GFP-expressing ES-derived germ cells occurred as colonies in the chimeric testes, where the seminiferous tubules were separated into green and non-green regions. When mature spermatozoa from green tubules were used for microinsemination, GFP-expressing offspring were efficiently obtained. Using a reverse study, we also obtained ES-derived progeny from GFP-negative ES cells in GFP-labeled host chimeras. Furthermore, we showed this approach could be accelerated by using round spermatids from the testes of 20-day-old chimeric mice. Thus, this technique allowed us to generate the ES cell-derived progeny even from the low contributed chimeric mice, which cannot produce ES-origin offspring by natural mating. genesis 43:34–42, 2005. © 2005 Wiley-Liss, Inc.

Get PDF (755K)