A non-invasive method for in situ quantification of subpopulation behaviour in mixed cell culture
A non-invasive method for in situ quantification of subpopulation behaviour in mixed cell culture
Ongoing advances in quantitative molecular- and cellular-biology highlight the need for correspondingly quantitative methods in tissue-biology, in which the presence and activity of specific cell-subpopulations can be assessed in situ. However, many experimental techniques disturb the natural tissue balance, making it difficult to draw realistic conclusions concerning in situ cell behaviour. In this study, we present a widely applicable and minimally invasive method which combines fluorescence cell labelling, retrospective image analysis and mathematical data processing to detect the presence and activity of cell subpopulations, using adhesion patterns in STRO-1 immunoselected human mesenchymal populations and the homogeneous osteoblast-like MG63 continuous cell line as an illustration.
Adhesion is considered on tissue culture plastic and fibronectin surfaces, using cell area as a readily obtainable and individual cell specific measure of spreading. The underlying statistical distributions of cell areas are investigated and mappings between distributions are examined using a combination of graphical and non-parametric statistical methods. We show that activity can be quantified in subpopulations as small as 1% by cell number, and outline behaviour of significant subpopulations in both STRO-1+/? fractions. This method has considerable potential to understand in situ cell behaviour and thus has wide applicability, for example in developmental biology and tissue engineering.
mathematical analysis, mesenchymal population, adhesion, image cytometry, tissue engineering
63-69
MacArthur, B.D.
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Tare, R.S.
587c9db4-e409-4e7c-a02a-677547ab724a
Please, C.P.
118dffe7-4b38-4787-a972-9feec535839e
Prescott, P.
cf0adfdd-989b-4f15-9e60-ef85eed817b2
Oreffo, R.C.
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22 February 2006
MacArthur, B.D.
2c0476e7-5d3e-4064-81bb-104e8e88bb6b
Tare, R.S.
587c9db4-e409-4e7c-a02a-677547ab724a
Please, C.P.
118dffe7-4b38-4787-a972-9feec535839e
Prescott, P.
cf0adfdd-989b-4f15-9e60-ef85eed817b2
Oreffo, R.C.
ff9fff72-6855-4d0f-bfb2-311d0e8f3778
MacArthur, B.D., Tare, R.S., Please, C.P., Prescott, P. and Oreffo, R.C.
(2006)
A non-invasive method for in situ quantification of subpopulation behaviour in mixed cell culture.
Journal of the Royal Society Interface, 3 (6), .
(doi:10.1098/rsif.2005.0080).
(PMID:16849218)
Abstract
Ongoing advances in quantitative molecular- and cellular-biology highlight the need for correspondingly quantitative methods in tissue-biology, in which the presence and activity of specific cell-subpopulations can be assessed in situ. However, many experimental techniques disturb the natural tissue balance, making it difficult to draw realistic conclusions concerning in situ cell behaviour. In this study, we present a widely applicable and minimally invasive method which combines fluorescence cell labelling, retrospective image analysis and mathematical data processing to detect the presence and activity of cell subpopulations, using adhesion patterns in STRO-1 immunoselected human mesenchymal populations and the homogeneous osteoblast-like MG63 continuous cell line as an illustration.
Adhesion is considered on tissue culture plastic and fibronectin surfaces, using cell area as a readily obtainable and individual cell specific measure of spreading. The underlying statistical distributions of cell areas are investigated and mappings between distributions are examined using a combination of graphical and non-parametric statistical methods. We show that activity can be quantified in subpopulations as small as 1% by cell number, and outline behaviour of significant subpopulations in both STRO-1+/? fractions. This method has considerable potential to understand in situ cell behaviour and thus has wide applicability, for example in developmental biology and tissue engineering.
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Published date: 22 February 2006
Keywords:
mathematical analysis, mesenchymal population, adhesion, image cytometry, tissue engineering
Organisations:
Statistics
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Local EPrints ID: 29272
URI: http://eprints.soton.ac.uk/id/eprint/29272
ISSN: 1742-5689
PURE UUID: 264b7473-4266-4621-a6a8-e559e87f1e1c
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Date deposited: 11 May 2006
Last modified: 16 Mar 2024 03:39
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C.P. Please
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