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Research Papers

Elasticity of Human Embryonic Stem Cells as Determined by Atomic Force Microscopy

[+] Author and Article Information
Robert Kiss1

Chemical Engineering, School of Engineering and Physical Sciences,  Heriot-Watt University, Edinburgh EH14 4AS, U.K.

Henry Bock, Nicholas A. Willoughby

Chemical Engineering, School of Engineering and Physical Sciences,  Heriot-Watt University, Edinburgh EH14 4AS, U.K.

Steve Pells, Paul De Sousa

MRC Centre for Regenerative Medicine, College of Medicine and Veterinary Medicine,  Edinburgh University, Edinburgh EH16 4SB, U.K.

Elisabetta Canetta2

BIONTHE (Bio- and Nano-Technologies for Health and Environment) Center, Division of Biotechnology and Forensic Sciences, School of Contemporary Sciences,  University of Abertay Dundee, Dundee DD1 1HG, U.K.

Ashok K. Adya

BIONTHE (Bio- and Nano-Technologies for Health and Environment) Center, Division of Biotechnology and Forensic Sciences, School of Contemporary Sciences,  University of Abertay Dundee, Dundee DD1 1HG, U.K.

Andrew J. Moore

Mechanical Engineering, School of Engineering and Physical Sciences,  Heriot-Watt University, Edinburgh EH14 4AS, U.K.

To be consistent with other publications in the field the standard deviation was used to indicate the “error” although the distribution in Fig. 3 is not Gaussian.

1

Corresponding author.

2

Present address: Cardiff School of Biosciences, Biomedical Sciences Building, Museum Avenue, Cardiff, CF10 3AX, U.K.

J Biomech Eng 133(10), 101009 (Nov 07, 2011) (9 pages) doi:10.1115/1.4005286 History: Received December 03, 2010; Accepted August 22, 2011; Published November 07, 2011; Online November 07, 2011

The expansive growth and differentiation potential of human embryonic stem cells (hESCs) make them a promising source of cells for regenerative medicine. However, this promise is off set by the propensity for spontaneous or uncontrolled differentiation to result in heterogeneous cell populations. Cell elasticity has recently been shown to characterize particular cell phenotypes, with undifferentiated and differentiated cells sometimes showing significant differences in their elasticities. In this study, we determined the Young’s modulus of hESCs by atomic force microscopy using a pyramidal tip. Using this method we are able to take point measurements of elasticity at multiple locations on a single cell, allowing local variations due to cell structure to be identified. We found considerable differences in the elasticity of the analyzed hESCs, reflected by a broad range of Young’s modulus (0.05-10 kPa). This surprisingly high variation suggests that elasticity could serve as the basis of a simple and efficient large scale purification/separation technique to discriminate subpopulations of hESCs.

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Copyright © 2011 by American Society of Mechanical Engineers
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Figures

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Figure 1

(a) One representative force-distance (F(d)) (dashed line) and force-indentation (F(δ)) (normal line) curve pair for Matrigel. Cantilever deflection is indicated by Δx. (b) The corresponding Young’s modulus versus indentation curve (E(δ)) obtained by piecewise fitting of Eq. 4.

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Figure 2

AFM image of Matrigel (a) and the corresponding force-indentation curves (b). Positions of recorded force curves are indicated by white spots on the AFM image and are numbered according to the order of the measurement.

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Figure 3

Probability distribution of Young’s moduli calculated at 61 different locations of Matrigel in the stable indentation region 0.1 μm ≤ δ ≤ 0.3 μm

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Figure 4

Probability distribution of Young’s moduli calculated at different locations of Fibroblast #1 (normal curve) and #2 (dashed curve) in the stable indentation region 0.1 μm ≤ δ ≤ 0.5 μm

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Figure 5

AFM images of RCM1 #1 (a), RCM1 #2 (b) and RCM1 #3 (c). Probability distributions of Young’s moduli (RCM1 #1 – dashed cyan line, RCM1 #2 – dashed violet line, RCM1 #3 – thick red line, RCM1 #4 – thick black line, RCM1 #5 – dashed yellow line and RCM1 #6 – thick green line) calculated in the stable indentation region 0.1 μm ≤ δ ≤ 0.5 μm (d). RCM1 #1 showed different morphology and a two orders of magnitude difference in its elasticity compared to RCM1 cells #2 and #3.

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Figure 6

AFM images of RH1 #1 (points 0-8) and RH1 #2 (points 9-21) (a) RH1 #3 (points 0-11) and RH1 #4 (points 12-24) (b). Probability distributions of Young’s moduli (RH1 #1 – thick green line, RH1 #2 – dashed cyan line, RH1 #3 – dashed violet line, RH1 #4 – thick red line, RH1 #5 – thick black line) calculated at different locations in the stable indentation region 0.1 μm ≤ δ ≤ 0.5 μm (c).

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Figure 7

AFM image and corresponding Young’s modulus versus indentation curves of Fibroblast cells #1 (a) and #2 (b).

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