Aggravation of cold-induced injury in Vero-B4 cells by RPMI 1640 medium

Identification of the responsible medium components

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Background: In modern biotechnology, there is a need for pausing cell lines by cold storage to adapt large-scale cell cultures to the variable demand for their products. We compared various cell culture media/solutions for cold storage of Vero-B4 kidney cells, a cell line widely used in biotechnology.
Results: Cold storage in RPMI 1640 medium, a recommended cell culture medium for Vero-B4 cells, surprisingly, strongly enhanced cold-induced cell injury in these cells in comparison to cold storage in Krebs-Henseleit buffer or other cell culture media (DMEM, L-15 and M199). Manufacturer, batch, medium supplements and the most likely components with concentrations outside the range of the other media/solutions (vitamin B12, inositol, biotin, p-aminobenzoic acid) did not cause this aggravation of cold-induced injury in RPMI 1640. However, a modified Krebs-Henseleit buffer with a low calcium concentration (0.42 mM), a high concentration of inorganic phosphate (5.6 mM), and glucose (11.1 mM; i.e. concentrations as in RPMI 1640) evoked a cell injury and loss of metabolic function corresponding to that observed in RPMI 1640. Deferoxamine improved cell survival and preserved metabolic function in modified Krebs-Henseleit buffer as well as in RPMI 1640. Similar Ca²⁺ and phosphate concentrations did not increase cold-induced cell injury in the kidney cell line LLC-PK1, porcine aortic endothelial cells or rat hepatocytes. However, more extreme conditions (Ca²⁺ was nominally absent and phosphate concentration raised to 25 mM as in the organ preservation solution University of Wisconsin solution) also increased cold-induced injury in rat hepatocytes and porcine aortic endothelial cells.
Conclusion: These data suggest that the combination of low calcium and high phosphate concentrations in the presence of glucose enhances cold-induced, iron-dependent injury drastically in Vero-B4 cells, and that a tendency for this pathomechanism also exists in other cell types.
PURL / DOI:
Lesezeichen:
Dokumententyp:
Wissenschaftliche Texte » Artikel, Aufsatz
Fakultät / Institut:
Medizinische Fakultät » Universitätsklinikum Essen » Institut für Physiologische Chemie
Medizinische Fakultät » Universitätsklinikum Essen » Klinik für Nephrologie
Dewey Dezimal-Klassifikation:
600 Technik, Medizin, angewandte Wissenschaften » 610 Medizin und Gesundheit
Stichwörter:
Calcium, Cell pausing, Cold storage, Hypothermia, Iron chelator, Phosphate, Preservation
Sprache:
Englisch
Kollektion / Status:
E-Publikationen / Dokument veröffentlicht
Dateien geändert am:
21.09.2018
Medientyp:
Text
Rechtliche Vermerke:
© 2012 Pless-Petig et al.; licensee BioMed Central Ltd.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Quelle:
BMC Biotechnology 2012, 12:73; Published: 10 October 2012