Fructose oligosaccharides as novel cryoprotectants for mammalian cells

PETER KILBRIDE; NELSON ROMANO; AYELEN HUGO; JULIE MENEGHEL; JEROME DELETTRE; JOHN MORRIS; ANDREA GÓMEZ-ZAVAGLIA; STEPHANIE PASSOT; FERNANDA FONSECA

Sevilla

Conferencia; 55th SLTB Scientific Conference; 2019

SLBT, Universidad de Sevilla

Successfulcryopreservation of mammalian somatic cells requires the use of cryoprotectants,among which dimethyl sulphoxide (DMSO) remains the most common. The cryoprotective abilities of DMSO were first recognized 60 yearsago (1). However, DMSO presents a cytotoxicity risk to cells, requiring preciseand careful use: it must be added to cell suspensions under chilled conditions,washed out or diluted post-thaw. The long-standing demand to develop newcryoprotectants which may be used instead of or in conjunction with lower dosesof DMSO remains unanswered. Fructose-based oligosaccharides(FOS), demonstrating cryo- and lyo-protective abilities on bacterial cells (2),appear as a potential alternative to DMSO for the cryopreservation of mammaliancells. In this study, we have examined the use of FOS solutions ascryoprotectants in a range of mammalian somatic cell types. Cryoprotection byFOS was observed with Chinese Hamster Ovarian cells, Jurkat (immortalized Tcell line), and CACO-2 (colorectal adherent) cells, with cell membraneintegrity, Alamar blue functionality, and proliferation of the cells observed.Disparate cell types were specifically chosen to determine if FOS could be usedmore generally as a mammalian somatic cryoprotectant and was not cell specific.The optimal concentration and composition of FOS solutions were determined. To better understand their cryoprotective mechanisms, therelationship between the degree of cryoprotection and some physical propertiesof the FOS solutions (nucleation temperature, glass transition temperature,osmolarity, viscosity) was also investigated.This work forms the basis of usingFOS as a cryoprotectant for mammalian cells, and this group of sugars may provea novel source for mammalian somatic cell cryopreservation. (1) Lovelock JE & Bishop MW (1959)Nature 16;183(4672):1394-5. (2) Romano N, Schebor C, Mobili P etal (2016) Food Res. Int. 90, 251-8.