Preparative Method for Obtaining Enucleated Mammalian-Cells

Wigler, M. H., Weinstein, I. B. (1975) Preparative Method for Obtaining Enucleated Mammalian-Cells. Biochemical and Biophysical Research Communications, 63 (3). pp. 669-674. ISSN 0006-291X

URL: http://www.ncbi.nlm.nih.gov/pubmed/1169063
DOI: 10.1016/S0006-291X(75)80436-0

Abstract

Mouse L cells can be enucleated in suspension by centrifugation in discontinuous Ficoll density gradients while in the presence of Cytochalasin B. Greater than 50% of the cytoplasts thus obtained attach to glass or plastic and undergo morphologic recovery within 2–4 hours of replating. Protein synthesis in cytoplasts undergoes a biphasic decay from an initial rate of approximately 50% of control nucleated cells. This method can yield up to 5 × 108 cytoplasts with consistently low levels of contamination by nucleated cells (less than 0.2%), and is well suited for obtaining quantitative amounts of cytoplasts or karyoplasts for physiologic or biochemical studies.

Item Type: Paper
Subjects: Investigative techniques and equipment
organs, tissues, organelles, cell types and functions > cell types and functions
CSHL Authors:
Communities: CSHL labs > Wigler lab
Depositing User: CSHL Librarian
Date Deposited: 06 Apr 2012 17:49
Last Modified: 02 May 2016 19:12
Related URLs:
URI: http://repository.cshl.edu/id/eprint/25955

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