Wound Microbiome Cryogenics

Polymicrobial infections contribute to persistence of non-healing wounds and exhibit complex dynamics not observed in culture. It was recently shown that patient-isolated wound slough could be transferred to recreate polymicrobial infection in a mouse wound model. Cryogenic archival is appreciated to enhance genomic and metagenomic studies, but its application for in vivo modeling is unknown. In the present study, effects of cryogenic preservation on colonization into mouse wounds were investigated through analysis of cell viability and microbiome community colonization dynamics. Freezing, either directly in liquid nitrogen or by first suspending in glycerol solution, did not reduce bacterial viability relative to fresh wound slough. Comparative analyses of microbiomes harvested from infected mouse wounds revealed a significant dependence on patient origin but not treatment type. Furthermore, overall bacterial load and the prior relative abundance of individual bacterial species in the patient wound were both found to significantly influence colonization on mice. Results describe characteristic properties of wound microbiome transfer from human to mouse, indicate resilience to cryogenic preservation and support the utility of cryogenic archival for a range of applications.