Mass isolation and culture of sea urchin micromeres |
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Authors: | Michael A Harkey Arthur H Whiteley |
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Institution: | (1) Department of Zoolology and Friday Harbor Laboratories, University of Washington, 98195 Seattle, Washington |
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Abstract: | Summary A procedure is described for large-scale isolation of micromeres from 16-cell stage sea urchin embryos. One to two grams of
>99% pure, viable micromeres (2.3 to 4.6 × 108 cells) are routinely isolated in a single preparation. In culture, these cells uniformly proceed through their normal development,
in synchrony with micromeres in whole embryos, ultimately differentiating typical larval skeletal structures. The attributes
of this procedure are: (a) the very early time of isolation of the cells, directly after the division that establishes the
cell line; (b) the large yield of cells; (c) the purity of the preparation of cell; and (d) their synchronous development
in culture through skeletogenesis. The procedure greatly aids in making sea urchin micromeres a favorable material for molecular
analysis of development.
This work was supported in part by the following grants from the National Institutes of Health: Grant HL-10312 to A.H.W.,
Grant GM-20784 to Helen R. Whiteley, Grant ES-02190 to N. Karle Mottet, M.D., and Training Grants ES-07032 and HD-00266. |
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Keywords: | micromere primary mesenchyme sea urchin isolation development |
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