Rs Biomarkers Job Force,” AustralianEthical ApprovalThe study was authorized by the nearby Ethics Committee with the Medical Faculty from the University of Leipzig (no. 351-1013122010).Conflict of InterestsProfessor H. Himmerich received speaker honorarium from AstraZeneca, Lilly, and Servier; consulting fees from BristolMyers Squibb; and chemical substances for study support from Lundbeck, AstraZeneca, Novartis, and Wyeth. All other authors reported no biomedical monetary interests or potential conflict of interests.Author’s ContributionH. Himmerich and S. Bartsch contributed equally to the paper.AcknowledgmentThe study was supported by the Claussen-Simon Foundation. The mentioned sponsor did not have any influence on study design, collection, evaluation, and interpretation of information; writing on the report; or the selection to submit the paper for publication.
Journal of Visualized Experiments Video ArticlejoveFormulations for Freeze-drying of Bacteria and Their Influence on Cell SurvivalPer Wessman , Sebastian H ansson , Klaus Leifer , Stefano Rubino1 two 1 1 2Department of Microbiology, Uppsala Biocenter, Swedish University of Agricultural Sciences Division of Engineering Sciences, Uppsala UniversityCorrespondence to: Stefano Rubino at stefano.BuyDiethyl (aminomethyl)phosphonate [email protected] URL: http://jove/video/4058 DOI: doi:10.3791/4058 Keywords and phrases: Microbiology, Problem 78, Cellular Biology, Molecular Biology, Biochemistry, Biophysics, Standard Protocols, Cell survival, sucrose, polysaccharides, cellulose, Ficoll, freeze-drying, Pseudomonas putida, Arthrobacter chlorophenolicus, cells, cell culture Date Published: 8/3/2013 Citation: Wessman, P., H ansson, S., Leifer, K., Rubino, S. Formulations for Freeze-drying of Bacteria and Their Influence on Cell Survival. J. Vis. Exp. (78), e4058, doi:10.3791/4058 (2013).AbstractCellular water might be removed to reversibly inactivate microorganisms to facilitate storage. A single such process of removal is freeze-drying, that is regarded as a gentle dehydration process. To facilitate cell survival for the duration of drying, the cells are typically formulated beforehand. The formulation forms a matrix that embeds the cells and protects them from a variety of dangerous stresses imposed around the cells throughout freezing and drying. We present right here a basic system to evaluate the survival rate of cells right after freeze-drying and we illustrate it by comparing the results obtained with four various formulations: the disaccharide sucrose, the sucrose derived polymer Ficoll PM400, and also the respective polysaccharides hydroxyethyl cellulose (HEC) and hydroxypropyl methyl cellulose (HPMC), on two strains of bacteria, P.4-bromo-2,6-dimethylpyridine Chemical name putida KT2440 and a.PMID:24624203 chlorophenolicus A6. Within this work we illustrate tips on how to prepare formulations for freeze-drying and the best way to investigate the mechanisms of cell survival following rehydration by characterizing the formulation employing of differential scanning calorimetry (DSC), surface tension measurements, X-ray analysis, and electron microscopy and relating these data to survival prices. The polymers had been selected to obtain a monomeric structure of the respective polysaccharide resembling sucrose to a varying degrees. Using this technique setup we showed that polymers can help cell survival as 1 efficiently as disaccharides if particular physical properties in the formulation are controlled .Video LinkThe video component of this short article may be discovered at http://jove/video/4058/Protocol1. Cultivation and harvest of P. putida1. Prepare a starter culture of Pseudomonas putida by inoculati.
