Technical Briefs

Development and Validation of a System for the Growth of Cells and Tissues Under Intermittent Hydrostatic Pressure

[+] Author and Article Information
Troy J. Eggum, Christopher J. Hunter

Department of Mechanical and Manufacturing Engineering, Centre for Bioengineering Research and Education, University of Calgary, Calgary, AB, T2N124, Canada

J Biomech Eng 130(6), 064501 (Oct 10, 2008) (6 pages) doi:10.1115/1.2979864 History: Received November 16, 2007; Revised April 08, 2008; Published October 10, 2008

Various cell populations have been shown to respond to hydrostatic pressure; however, many of the culture systems suffer from shortcomings in design or methodology. Of particular interest to us is the potential role of pressure and other environmental factors in modulating stem cell behavior in intervertebral disk repair. A system was developed for the growth of cells and tissues under intermittent hydrostatic pressure. The system was validated with NIH 3T3 fibroblasts for sterilizability and cytotoxicity. Further experiments were conducted with canine mesenchymal stem cells under various levels of pressure, oxygen, glucose, and conditioned medium. The culture system showed no cytotoxicity and was able to demonstrate that the proliferation and metabolism of mesenchymal stem cells are sensitive to medium glucose and oxygen concentration and hydrostatic pressure. The cells exposed to hydrostatic pressure differed in their morphology from nonexposed cells. The system is capable of supporting long-term cell culture and examining the role of mechanical and environmental stimulation in vivo.

Copyright © 2008 by American Society of Mechanical Engineers
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Figure 1

The hydrostatic pressure bioreactor, indicating the sequence of assembly. (a) The baseplate, (b) the acrylic multidish, (c) the neoprene pressure transfer membrane, (d) the manifold, (e) the lid, and (f) the complete system attached to the pump system.

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Figure 2

Plot of fluid volume versus pressure. The total fluid volume is 66ml; thus, the system compressibility is approximately 0.2%.

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Figure 3

Photomicrograph of wells after sterility testing. No evidence of bacterial contamination was found; the medium was clear, and no organisms could be found upon microscopic examination.

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Figure 4

Photomicrograph of cells grown in 5% oxygen and under atmospheric (a) and (c) and 1MPa pressure for 1h (b) and (d), noting the rounded morphology of the loaded cells. No evidence of necrotic or apoptotic cells was found.

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Figure 5

Main effects’ plot for MTT and pH, indicating the direction of change for each factor examined. Hydrostatic pressure (HSP); mesenchymal stem cell conditioned medium (MSC) (control); Noto: notochordal cell conditioned medium. The stars indicate significant effects.

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Figure 6

Main effects’ plots for residual glucose and lactate. Hydrostatic pressure (HSP); mesenchymal stem cell conditioned medium (MSC) (control); notochordal cell conditioned medium (Noto). The stars indicate significant effects.

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Figure 7

Main effects’ plots for glutamine and glutamate. Hydrostatic pressure (HSP); mesenchymal stem cell conditioned medium (MSC) (control); notochordal cell conditioned medium (Noto). The stars indicate significant effects.




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