A rapid, cost-effective method for counting human embryonic stem cell numbers as clumps

Prowse, ABJ, Wolvetang, EJ and Gray, PP (2009) A rapid, cost-effective method for counting human embryonic stem cell numbers as clumps. Biotechniques, 47 1: 599-606. doi:10.2144/000113151


Author Prowse, ABJ
Wolvetang, EJ
Gray, PP
Title A rapid, cost-effective method for counting human embryonic stem cell numbers as clumps
Journal name Biotechniques   Check publisher's open access policy
ISSN 0736-6205
Publication date 2009-07-01
Year available 2009
Sub-type Article (original research)
DOI 10.2144/000113151
Open Access Status
Volume 47
Issue 1
Start page 599
End page 606
Total pages 5
Editor Nathan Blow
Place of publication New York , U. S. A.
Publisher Informa Healthcare
Language eng
Subject C1
Abstract Enumeration of human embryonic stem cell (hESC) numbers through single cell digestion can be time consuming especially in high-throughput or multifactorial analysis containing 50+ samples. We have developed a reproducible, cost-effective method of counting hESCs in clumps circumventing the need to manually dissociate each sample to single cells. The method is based on the DNA binding capacity of propidium iodide (PI) and subsequent fluorescent signal detection. Standard curves generated for cell numbers versus PI fluorescence as single cells or clumps showed an almost identical relationship in the lines of best fit. The reproducibility of the assay was first demonstrated by seeding hESC clumps at specific cell densities ranging 0.05-2 x 10(5) cells/well and then secondly by using the assay to count cell numbers after different growth conditions. Validation tests showed that consistent seeding densities are important in maintaining undifferentiated hESC culture and that the assay can be used to estimate relative cell numbers and growth curves with high accuracy.
Keyword human embryonic stem cells
DIFFERENTIATION
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: 2010 Higher Education Research Data Collection
Australian Institute for Bioengineering and Nanotechnology Publications
 
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Citation counts: TR Web of Science Citation Count  Cited 8 times in Thomson Reuters Web of Science Article | Citations
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Created: Thu, 03 Sep 2009, 17:48:33 EST by Mr Andrew Martlew on behalf of Aust Institute for Bioengineering & Nanotechnology