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  • A Guide for Ex Vivo Handling and Storage of Stool Samples Intended for Fecal Microbiota Transplantation.

A Guide for Ex Vivo Handling and Storage of Stool Samples Intended for Fecal Microbiota Transplantation.

Scientific reports (2019-06-22)
Sebastian D Burz, Anne-Laure Abraham, Fernanda Fonseca, Olivier David, Audrey Chapron, Fabienne Béguet-Crespel, Stéphanie Cénard, Karine Le Roux, Orlane Patrascu, Florence Levenez, Carole Schwintner, Hervé M Blottière, Christel Béra-Maillet, Patricia Lepage, Joël Doré, Catherine Juste
ABSTRACT

Owing to the growing recognition of the gut microbiota as a main partner of human health, we are expecting that the number of indications for fecal microbiota transplantation (FMT) will increase. Thus, there is an urgent need for standardization of the entire process of fecal transplant production. This study provides a complete standardized procedure to prepare and store live and ready-to-use transplants that meet the standard requirements of good practices to applied use in pharmaceutical industry. We show that, if time before transformation to transplants would exceed 24 hours, fresh samples should not be exposed to temperatures above 20 °C, and refrigeration at 4 °C can be a safe solution. Oxygen-free atmosphere was not necessary and simply removing air above collected samples was sufficient to preserve viability. Transplants prepared in maltodextrin-trehalose solutions, stored in a -80 °C standard freezer and then rapidly thawed at 37 °C, retained the best revivification potential as  proven by 16S rRNA profiles, metabolomic fingerprints, and flow cytometry assays over a 3-month observation period. Maltodextrin-trehalose containing cryoprotectants were also efficient in preserving viability of lyophilized transplants, either in their crude or purified form, an option that can be attractive for fecal transplant biobanking and oral formulation.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
(+)-Sodium L-ascorbate, crystalline, ≥98%
Sigma-Aldrich
D-(+)-Maltose monohydrate, from potato, ≥99%
Sigma-Aldrich
L-Cysteine hydrochloride monohydrate, reagent grade, ≥98% (TLC)
Millipore
D-(+)-Trehalose dihydrate, ≥99.0%, suitable for microbiology
Sigma-Aldrich
Hemin, from bovine, ≥90%
SAFC
HEPES