Mammalian cell survival and processing in supercritical CO2

Ginty, Patrick J., Howard, Daniel, Rose, Felicity R.A.J., Whitaker, Martin J., Barry, John J.A., Tighe, Patrick, Mutch, Stacey R., Serhatkulu, Gulay, Oreffo, Richard O.C., Howdle, Steven M. and Shakesheff, Kevin M. (2006) Mammalian cell survival and processing in supercritical CO2. Proceedings of the National Academy of Sciences of the United States of America, 103 (19), 7426-7431. (doi:10.1073/pnas.0508895103).

Abstract

We demonstrate that mammalian cells can survive for 5 min within high-pressure CO(2)(.) Cell survival was investigated by exposing a range of mammalian cell types to supercritical CO(2) (scCO(2)) (35 degrees C, 74 bar; 1 bar = 100 kPa) for increasing exposure and depressurization times. The myoblastic C2C12 cell line, 3T3 fibroblasts, chondrocytes, and hepatocytes all displayed appreciable but varying metabolic activity with exposure times up to 1 min. With depressurization times of 4 min, cell population metabolic activity was >/=70% of the control population. Based on survival data, we developed a single-step scCO(2) technique for the rapid production of biodegradable poly(dl-lactic acid) scaffolds containing mammalian cells. By using optimum cell-survival conditions, scCO(2) was used to process poly(dl-lactic acid) containing a cell suspension, and, upon pressure release, a polymer sponge containing viable mammalian cells was formed. Cell functionality was demonstrated by retention of an osteogenic response to bone morphogenetic protein-2 in C2C12 cells. A gene microarray analysis showed no statistically significant changes in gene expression across 4,418 genes by a single-class t test. A significance analysis of microarrays revealed only eight genes that were down-regulated based on a delta value of 1.0125 and a false detection rate of 0.

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Contributors

Author: Richard O.C. Oreffo

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