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Echinobase
ECB-ART-43285
Methods Mol Biol 2014 Jan 01;1128:187-96. doi: 10.1007/978-1-62703-974-1_12.
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Isolating specific embryonic cells of the sea urchin by FACS.

Juliano C , Swartz SZ , Wessel G .


Abstract
Isolating cells based on specific gene expression enables a focused biochemical and molecular analysis. While cultured cells and hematopoietic cells, for example, are routinely isolated by fluorescence activated cell sorting (FACS), early embryonic cells are a relatively untapped source for FACS applications often because the embryos of many animals are quite limiting. Furthermore, many applications require genetic model organisms in which cells can be labeled by fluorescent transgenes, or antibodies against cell surface antigens. Here we define conditions in the sea urchin embryo for isolation of embryonic cells based on expression of specific proteins. We use the sea urchin embryo for which a nearly unlimited supply of embryonic cells is available and demonstrate the conditions for separation of the embryo into single cells, fixation of the cells for antibody penetration into the cells, and conditions for FACS of a rare cell type in the embryo. This protocol may be adapted for analysis of mRNA, chromatin, protein, or carbohydrates and depends only on the probe availability for the cell of interest. We anticipate that this protocol will be broadly applicable to embryos of other species.

PubMed ID: 24567215
PMC ID: PMC4108900
Article link: Methods Mol Biol
Grant support: [+]

Genes referenced: LOC100887844 LOC115919910
Antibodies: LOC373239 Ab1 ddx4 Ab1 ddx4 Ab2 nanos2l Ab1 tgml Ab1

References [+] :
Bruskin, Accumulation in embryogenesis of five mRNAs enriched in the ectoderm of the sea urchin pluteus. 1981, Pubmed, Echinobase