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A new method for isolating primary mesenchyme cells of the sea urchin embryo. Panning on wheat germ agglutinin-coated dishes.

Publication ,  Journal Article
Ettensohn, CA; McClay, DR
Published in: Experimental cell research
February 1987

This paper describes a rapid and efficient way to isolate primary mesenchyme cells (PMCs) of the sea urchin embryo. The procedure involves three simple steps: Dissociation of mesenchyme blastulae in calcium-free artificial seawater. Incubation of the resulting cell suspension on dishes that have been coated with wheat germ agglutinin (WGA), to which the PMCs adhere more firmly than do other cell types. Gentle rinsing of the dishes to remove loosely attached cells, followed by more vigorous rinsing to remove PMCs. This panning procedure has been applied to embryos of three species of sea urchins, Lytechinus variegatus, L. pictus and Arbacia punctulata, and yields populations of PMCs that are 95-99% pure as determined by the proportion of cells that stain with fluorescein isothiocynate (FITC)-WGA and with a monoclonal antibody that binds specifically to PMCs. The yield of PMCs is 4-5 X 10(6) cells/100-mm dish, or 1-2 X 10(7) PMCs/ml of packed embryos. The principal advantages of this procedure are that it can be carried out rapidly and simply, and it yields pure populations of PMCs.

Duke Scholars

Published In

Experimental cell research

DOI

EISSN

1090-2422

ISSN

0014-4827

Publication Date

February 1987

Volume

168

Issue

2

Start / End Page

431 / 438

Related Subject Headings

  • Wheat Germ Agglutinins
  • Sea Urchins
  • Fluoresceins
  • Fluorescein-5-isothiocyanate
  • Embryo, Nonmammalian
  • Cell Separation
  • Blastocyst
  • Biochemistry & Molecular Biology
  • Antibodies, Monoclonal
  • Animals
 

Citation

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Ettensohn, C. A., & McClay, D. R. (1987). A new method for isolating primary mesenchyme cells of the sea urchin embryo. Panning on wheat germ agglutinin-coated dishes. Experimental Cell Research, 168(2), 431–438. https://doi.org/10.1016/0014-4827(87)90015-2
Ettensohn, C. A., and D. R. McClay. “A new method for isolating primary mesenchyme cells of the sea urchin embryo. Panning on wheat germ agglutinin-coated dishes.Experimental Cell Research 168, no. 2 (February 1987): 431–38. https://doi.org/10.1016/0014-4827(87)90015-2.
Ettensohn, C. A., and D. R. McClay. “A new method for isolating primary mesenchyme cells of the sea urchin embryo. Panning on wheat germ agglutinin-coated dishes.Experimental Cell Research, vol. 168, no. 2, Feb. 1987, pp. 431–38. Epmc, doi:10.1016/0014-4827(87)90015-2.
Journal cover image

Published In

Experimental cell research

DOI

EISSN

1090-2422

ISSN

0014-4827

Publication Date

February 1987

Volume

168

Issue

2

Start / End Page

431 / 438

Related Subject Headings

  • Wheat Germ Agglutinins
  • Sea Urchins
  • Fluoresceins
  • Fluorescein-5-isothiocyanate
  • Embryo, Nonmammalian
  • Cell Separation
  • Blastocyst
  • Biochemistry & Molecular Biology
  • Antibodies, Monoclonal
  • Animals