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Formulations for freeze-drying of bacteria and their influence on cell survival
Swedish Univ Agr Sci, Uppsala Bioctr, Dept Microbiol, S-90183 Umea, Sweden.
Swedish Univ Agr Sci, Uppsala Bioctr, Dept Microbiol, S-90183 Umea, Sweden.
Uppsala University, Disciplinary Domain of Science and Technology, Technology, Department of Engineering Sciences, Applied Materials Sciences. (Electron Microscopy and Nano-Engineering)
Uppsala University, Disciplinary Domain of Science and Technology, Technology, Department of Engineering Sciences, Applied Materials Sciences.
2013 (English)In: Journal of Visualized Experiments, ISSN 1940-087X, E-ISSN 1940-087X, no 78, e4058Article in journal (Refereed) Published
Abstract [en]

Cellular water can be removed to reversibly inactivate microorganisms to facilitate storage. One such method of removal is freeze-drying, which is considered a gentle dehydration method. To facilitate cell survival during drying, the cells are often formulated beforehand. The formulation forms a matrix that embeds the cells and protects them from various harmful stresses imposed on the cells during freezing and drying. We present here a general method to evaluate the survival rate of cells after freeze-drying and we illustrate it by comparing the results obtained with four different formulations: the disaccharide sucrose, the sucrose derived polymer Ficoll PM400, and the respective polysaccharides hydroxyethyl cellulose (HEC) and hydroxypropyl methyl cellulose (HPMC), on two strains of bacteria, P. putida KT2440 and A. chlorophenolicus A6. In this work we illustrate how to prepare formulations for freeze-drying and how to investigate the mechanisms of cell survival after rehydration by characterizing the formulation using of differential scanning calorimetry (DSC), surface tension measurements, X-ray analysis, and electron microscopy and relating those data to survival rates. The polymers were chosen to get a monomeric structure of the respective polysaccharide resembling sucrose to a varying degrees. Using this method setup we showed that polymers can support cell survival as effectively as disaccharides if certain physical properties of the formulation are controlled.

Place, publisher, year, edition, pages
2013. no 78, e4058
National Category
Medical and Health Sciences Engineering and Technology
Research subject
Engineering Science with specialization in Materials Science
Identifiers
URN: urn:nbn:se:uu:diva-223090DOI: 10.3791/4058ISI: 000209228000001PubMedID: 23963171OAI: oai:DiVA.org:uu-223090DiVA: diva2:712757
Funder
Mistra - The Swedish Foundation for Strategic Environmental ResearchEU, FP7, Seventh Framework Programme, 211684
Available from: 2014-04-16 Created: 2014-04-16 Last updated: 2017-12-05Bibliographically approved

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Leifer, KlausRubino, Stefano

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