title: Preparation and preservation of viable Akkermansia muciniphila cells for therapeutic interventions.
abstract: The anaerobic gut bacterium Akkermansia muciniphila is a well-characterised member of the mucosal microbiota and has shown to be a gut symbiont in human. A. muciniphila has been negatively associated with obesity and its associated metabolic disorders in various human cohorts while treatment with A. muciniphila cells reversed highfat diet-induced obesity and its associated metabolic disorders in mouse models. Therefore, administration of A. muciniphila has been suggested as a possible new therapeutic treatment for these omnipresent diseases. Here we describe a potentially scalable workflow for the preparation and preservation of high numbers of viable cells of A. muciniphila obtained from 1 l laboratory scale growth under strict anaerobic conditions for therapeutic interventions. This resulted in viable A. muciniphila cells with high yields and very high stability, with up to 97.9±4.5% survival for a time period of 1 year at -80 °C in glycerol-amended medium. Moreover, various quality assessment and control procedures were developed to ensure the use of viable cells of A. muciniphila. Several microscopic, culturing, and molecular approaches were applied to monitor the presence, abundance and recovery of A. muciniphila before, during, and after its administration to high-fat treated mice. We show that viable A. muciniphila cells can be recovered from caecal and colon content (up to 1×1010cells/g), testifying for the efficiency of the described workflow.
authors.affiliation: 1 Laboratory of Microbiology, Wageningen University, Dreijenplein 10, 6703 HB Wageningen, the Netherlands.
authors.forename: J P
authors.initials: JP
authors.lastname: Ouwerkerk
authors.affiliation: 1 Laboratory of Microbiology, Wageningen University, Dreijenplein 10, 6703 HB Wageningen, the Netherlands.
authors.forename: S
authors.initials: S
authors.lastname: Aalvink
authors.affiliation: 1 Laboratory of Microbiology, Wageningen University, Dreijenplein 10, 6703 HB Wageningen, the Netherlands.
authors.forename: C
authors.initials: C
authors.lastname: Belzer
authors.affiliation: 1 Laboratory of Microbiology, Wageningen University, Dreijenplein 10, 6703 HB Wageningen, the Netherlands.|2 Department of Veterinary Biosciences, University of Helsinki, P.O. Box 66, 00014 Helsinki, Finland.|3 Department of Bacteriology and Immunology, and Research Programs Unit Immunology, University of Helsinki, P.O. Box 21, 00014 Helsinki, Finland.
authors.forename: W M
authors.initials: WM
authors.lastname: De Vos
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D004195:Disease Models, Animal; D000069196:Gastrointestinal Microbiome; D006801:Humans; D007413:Intestinal Mucosa / Q000382:microbiology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008820:Mice, Obese; D009765:Obesity / Q000628:therapy*; D019936:Probiotics / Q000008:administration & dosage* / Q000627:therapeutic use*; D061327:Verrucomicrobia / Q000166:cytology / Q000254:growth & development*
keywords: delivery; laboratory scale production; stability; therapeutic microbe; viability