From fresh to frozen: evaluating the feasibility of unprocessed raw stool banking for scalable FMT production
Fedja Farowski, Anastasia Tsakmaklis, Nina Fischer, Lena M. Biehl, Julia Heger, Klaus Schomäcker, Maria J. G. T. VehreschildBackground:
Manufacturing of fecal microbiota transplantation (FMT) products from fresh stool donations poses significant logistical, safety, and scalability challenges.
Objectives:
To evaluate the technical feasibility of freezing unprocessed, raw stool donations prior to manufacturing to improve process control and efficiency.
Design:
A paired study design compared the immediate processing of fresh stool with the delayed processing of raw stool stored at −80°C for 1–48 weeks.
Methods:
Stool donations from three donors were divided into two aliquots and processed immediately fresh or frozen using the identical protocol. Key quality attributes, including bacterial viability (fluorescent staining) and microbial community structure (16S rRNA gene sequencing), were compared.
Results:
Preparations from frozen material showed a significant reduction in viable cells compared to their fresh counterparts (mean 2.2 vs 6.5 × 10
10
cells/mL;
Conclusion:
The manufacturing of FMT products from unprocessed, cryopreserved stool is a standardized and logistically superior method. This approach yields preparations that, despite a predictable reduction in viability, retain high potency and a robustly preserved microbial community structure, facilitating the standardization and scalability required for the advancement of FMT.