Micalis Institute, INRA, AgroParisTech, Université Paris-Saclay, 78350, Jouy-en-Josas, France.
Université Paris-Saclay, MetaGenoPolis, INRA, 78350, Jouy-en-Josas, France.
Sci Rep. 2019 Jun 20;9(1):8897. doi: 10.1038/s41598-019-45173-4.
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.
由于人们越来越认识到肠道微生物群是人类健康的主要伙伴，我们预计粪便微生物群移植（FMT）的适应证数量将会增加。因此，迫切需要规范粪便移植生产的整个过程。本研究提供了一种完整的标准化程序，用于制备和储存符合良好生产规范标准要求的活菌和即用型移植体，以应用于制药工业。我们表明，如果转化为移植体的时间超过 24 小时，新鲜样本不应暴露在 20°C 以上的温度下，4°C 的冷藏可以是一种安全的解决方案。无氧环境不是必需的，只需将收集样本上方的空气抽出即可保持活力。在麦芽糖糊精-海藻糖溶液中制备的移植体，在-80°C 的标准冷冻器中储存，然后在 37°C 下迅速解冻，通过 16S rRNA 图谱、代谢组学指纹图谱和流式细胞术检测，在 3 个月的观察期内保留了最佳的复活潜力。含有冷冻保护剂的麦芽糖糊精-海藻糖在冻干移植体的存活方面也非常有效，无论是粗提物还是纯化形式，这对于粪便移植生物库和口服制剂来说都是一个有吸引力的选择。
