ATTEMPTS TO CULTURE ISOLATED URODELAN BLASTOMERES THAT CONTINUE TO DIVIDE WITHOUT AGGREGATING*

Authors

  • MAMORU KUSA,

    1. Laboratory of Developmental Biology, Faculty of Science, Yamagata University, Yamagata 990 and Department of Biology, Faculty of Science, Kanazawa University, Kanazawa 920, Japan
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  • HIDEO SATO

    1. Laboratory of Developmental Biology, Faculty of Science, Yamagata University, Yamagata 990 and Department of Biology, Faculty of Science, Kanazawa University, Kanazawa 920, Japan
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  • *

    This paper is dedicated to Prof. Emer. Katsuma Dan for celebration of his 70th birthday.

Abstract

The blastomeres isolated from urodelan blastulae continued to divide without aggregating of daughter cells when inoculated with Ca2+-free neutral Holtfreter solution into glass culture dishes coated with agar. When standard Holtfreter solution with pH 8.2 was used as a culture medium, Ca2+ content from 1/40 to 1/20 of the original strength was essential for the purpose of the present observations; other divalent cations such as Mg2+, Ba2+ or Mn2+ replaced Ca2+. Under these experimental conditions, cell pedigrees were obtained during the incubation period. The greatest number of cell divisions so far observed in vitro was 8 for Hynobius lichenatus, and 9 for Cynops pyrrhogaster. Some related observations on the behavior of isolated blastomeres are also presented.

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