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Automated mechanical passaging: a novel and efficient method for human embryonic stem cell expansion.

TitleAutomated mechanical passaging: a novel and efficient method for human embryonic stem cell expansion.
Publication TypeJournal Article
Year of Publication2006
AuthorsJoannides A, Fiore-Heriche C, Westmore K, Caldwell M, Compston A, Allen ND, Chandran S
JournalStem Cells
Volume24
Issue2
Pagination230-5
Date Published2006 Feb
ISSN1066-5099
KeywordsAnimals, Automation, Biological Markers, Cell Culture Techniques, Cell Line, Embryo, Mammalian, Humans, Mice, Pluripotent Stem Cells, Stem Cells, Time Factors
Abstract

There is a need for more standardized methods of maintenance and propagation of human embryonic stem cell (hESC) cultures. Enzymatic passaging currently represents the most widely used method for expansion of hESCs. Although rapid and straightforward, this technique results in variable-sized cell clusters and significant cellular trauma, which may apply selective pressure in long-term culture. Mechanical passaging has the potential advantages of defined colony fragment sizes, reduced cellular trauma, and the possibility of selecting undifferentiated colonies for transfer. However, manual dissection of individual colonies is a prohibitively time-consuming process unsuitable for maintaining large numbers of hESCs without the use of additional chemical means. In this study we report an efficient automated method for mechanically passaging hESCs. We have used this method exclusively to maintain hESCs in long-term undifferentiated culture without the use of enzymatic digestion for longer than 100 days. This automated technique can thus be used routinely to culture hESCs in the laboratory.

DOI10.1634/stemcells.2005-0243
Alternate JournalStem Cells
PubMed ID16510428
Grant ListG0300300 / / Medical Research Council / United Kingdom
Publication institute
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