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Mammalian cell survival and processing in supercritical CO(2).

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.

SUBMITTER: Ginty PJ

PROVIDER: S-EPMC1464355 | biostudies-literature | 2006 May

REPOSITORIES: biostudies-literature

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Mammalian cell survival and processing in supercritical CO(2).

Ginty Patrick J PJ Howard Daniel D Rose Felicity R A J FR Whitaker Martin J MJ Barry John J A JJ Tighe Patrick P Mutch Stacey R SR Serhatkulu Gulay G Oreffo Richard O C RO Howdle Steven M SM Shakesheff Kevin M KM

Proceedings of the National Academy of Sciences of the United States of America 20060501 19

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 ...[more]

PMID: 16651535

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Project description:Sugarcane juice is a nutritious and energetic drink. For its processing, the use of supercritical carbon dioxide (SC-CO2) technology as an intervention potentially capable of rendering a high quality product can be considered. This study evaluated the combined effect of SC-CO2 and mild temperatures, primarily aiming for the reduction of endogenous microorganisms and enzymes in sugarcane juice (pH~5.5). Pressures (P) ranging from 74 to 351 bar, temperatures (T) between 33 and 67 °C, and holding times (t) between 20 and 70 min were tested in a central composite rotational design. Seventeen trials were performed, comprising three replicates at the central points. Counts of aerobic mesophiles, molds and yeasts, lactic acid bacteria and coliforms at 45 °C, determination of polyphenol oxidase (PPO) and peroxidase (POD) activities, and measurement of color parameters in freshly extracted and processed juice's samples were carried out. The pH of fresh and processed juice varied between 4.6 and 6.0, and between 4.6 and 6.3, respectively. The number of decimal reductions achieved in mesophiles, molds and yeasts, lactic acid bacteria and coliforms varied between 0.1 and 3.9, 2.1 and 4.1, 0.0 and 2.1, and 0.3 to 2.5, respectively. The percentages of PPO reduction ranged from 3.51% to 64.18%. Regarding the POD, reductions between 0.27% and 41.42% were obtained. Color variations between fresh and processed samples varied between 2.0 and 12.3. As for mesophiles, molds and yeasts reduction, and soluble solids variation, none of the variables or their interactions were significant. In terms of polyphenol oxidase (PPO) reduction, only t was significant; however, T, t, and the interaction between them significantly affected the peroxidase (POD) reduction. In regards to pH variation, P, and the interaction between T and t were significant. P, T, t, and the interaction between T and t played a significant effect on color. The combination of mild temperatures and SC-CO2 can be potentially used for cane juice preservation.

Dataset Information

Mammalian cell survival and processing in supercritical CO(2).

Publications

Mammalian cell survival and processing in supercritical CO(2).

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