ABSTRACT

INTRODUCTION: FIVE BASIC METHODS OF VITRIFICATION OF CELLS It is well established that storage of cells in a liquid milieu leads to degradation processes, and to eventual loss of cell viability. At the same time, intracellular ice is lethal for the majority of cells.1 Thus, the only stage in which the cells can be stabilized would be a solid-like phase in which intracellular ice crystals are not formed, or have not grown to the ‘critical lethal size’. Such a vitreous (‘glassy’) state has elevated viscosity (in the range of 1012 Pa.s at the glass transition temperature Tg and up to 10

13.5 Pa.s at the so called ‘strain point),2 so processes of chemical and physical degradation are essentially stopped for the duration of experiments or storage. We can distinguish five basic methods to achieve intracellular vitrification; all of which lead to a drastic decrease of the water activity.3