Effects of dimethylsulfoxide and the deep-freezing process on the infectivity, motility, and ultrastructure of Trypanosoma cruzi

The effects of dimethylsulfoxide (DMSO, final concentration 5%) and the deep-freezing process on the infectivity (ID50), motility, and ultrastructure of nontreated and DMSO-treated Trypanosoma cruzi suspensions (PSG-3 buffer with 10% horse serum) were investigated prior to and after cryopreservation in liquid nitrogen. DMSO equilibration caused distinct suppression of motility and characteristic, fine structural alterations in numerous organelles, such as myelin-like structures in the cytoplasm and/or inside the mitochondrial apparatus, enlargement of the perinuclear space, endoplasmic reticulum, and mitochondrial cristae, as well as condensation of the kinetoplast with loss of its lamellar structure. There was no evidence of loss of infectivity in DMSO-treated parasites. DMSO-treated and deep-frozen organisms showed, however, very similar fine structural alterations, although damage occurring during freezing and thawing was more pronounced. Apart from the frequently enlarged kinetoplast and the loosening of its mitochondrial matrix, numerous trypanosomes revealed total disintegration of the kinetoplast-mitochondrion complex with loss of its whole matrix. Deep-frozen trypanosomes were significantly less infective to mice than nontreated organisms, and their motility was strongly suppressed. These results suggest that cryopreservation and thawing of T. cruzi may lead to severe damage of the mitochondrial apparatus and thus to heavy disorders of metabolic function, exhaustion of the metabolic pool, and finally, to death of such damaged trypanosomes, despite the use of DMSO as a cryoprotective agent.