Clonal Isolation and Plating Efficiency
Bernice M Martin, Suffolk University, Boston, Massachusetts, USA
Published online: December 2011
Viable cultures of cells that were initiated from single cells can be obtained. However, both the isolation of the single
cell of interest and the ability to promote proliferation of this cell present challenges. Selection of the single cell can
be achieved in a number of ways. Among the methods discussed are a direct approach (cloning cylinders or discs), a theoretical
approach (limiting dilution) and isolation using cell sorters. Any of these approaches can be combined with a pre‐seeding
technique that will enrich the population of the desired cells in the culture. A number of techniques to address the second
challenge, cell proliferation, are also addressed. These include the use of culture ware with a small growth area, increasing
the likelihood of cell attachment, the enrichment of the culture medium and the use of feeder layer cells. Because cells differ
widely in their characteristics, a combination of techniques can be used.
Cloned cells in culture are derived from a single cell.
Each of the cells in a clone should have identical characteristics.
Selection of cells for cloning may be aided by fluorescent molecules and FACS.
The survival and proliferation of the cells that form clones require special conditions.
Keywords: adherent cells; single‐cell isolation; single‐cell survival; cloning cylinders and discs; FACS
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Aside from journal articles, books and monographs, trade journals and trusted vendors are excellent sources of current advancements and background information. Examples of such are: Biotechniques.com: http://www.biotechniques.com
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Sigma‐Aldrich Informational Publications: http://www.sigma‐aldrich.com or http://www.sigma‐aldrich.com/techservice