Cell and Tissue Research

, Volume 350, Issue 2, pp 251–260 | Cite as

Fluorescent transgenic mice suitable for multi-color aggregation chimera studies

  • Masato OhtsukaEmail author
  • Hiromi Miura
  • Channabasavaiah B. Gurumurthy
  • Minoru Kimura
  • Hidetoshi Inoko
  • Shinichi Yoshimura
  • Masahiro Sato
Regular Article


We recently reported a novel method of mouse transgenesis called Pronuclear Injection-based Targeted Transgenisis (PITT) using which a series of fluorescent transgenic (Tg) mice lines were generated. These lines, unlike those generated using conventional random integration methods, express the transgenes faithfully and reproducibly generation after generation. Because of this superior nature, these lines are ideal for the generation of multi-colored aggregation chimeras that can be used to study cell–cell interactions and lineage analyses in living embryos/organs, where the transgenes can be detected and the clonal origin of a given cell population easily traced by its distinct fluorescence. In this study, to verify if Tg fluorescent mice generated through PITT were suitable for such applications, we sought to generate chimeric blastocysts and chimeric-Tg mice by aggregating two- or three-colored 8-cell embryos. Our analyses using these models led to the following observations. First, we noticed that cell mixing was infrequent during the stages of morula to early blastocyst. Second, chimeric fetuses obtained after aggregation of the two-colored 8-cell embryos exhibited uniform cell mixing. And third, in the organs of adult chimeric mice, the mode of cell distribution could be either clonal or polyclonal, as previously pointed out by others. Implications of our novel and improved Tg-chimeric mice approach for clonal cell lineage and developmental studies are discussed.


Fluorescent mouse Aggregation chimera Preimplantation embryos Mice Clonality 



We thank T. Sasou, M. Ando, N. Motosugi and S. Ogiwara for technical assistance. We are grateful to Manju George for critical reading of the manuscript and for English corrections. This work was supported by Grant-in-Aid for Young Scientists (B) [20700368] from the Ministry of Education, Culture, Sports, Science and Technology (MEXT); and by the Research and Study Program of the Tokai University Educational System General Research Organization (2008) to M.O.


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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Masato Ohtsuka
    • 1
    Email author
  • Hiromi Miura
    • 1
  • Channabasavaiah B. Gurumurthy
    • 2
  • Minoru Kimura
    • 1
  • Hidetoshi Inoko
    • 1
  • Shinichi Yoshimura
    • 1
  • Masahiro Sato
    • 3
  1. 1.Department of Molecular Life Science, Division of Basic Medical Science and Molecular MedicineTokai University School of MedicineIseharaJapan
  2. 2.Department of Genetics, Cell Biology and AnatomyUniversity of Nebraska Medical CenterOmahaUSA
  3. 3.Section of Gene Expression Regulation, Frontier Science Research CenterKagoshima UniversityKagoshimaJapan

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