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It means that the border between that Zone II, where vitrification is achievable with the Zone I, where successfully vitrification is impossible at any “reasonable speed” of cooling and warming largely depends on the rate of that cooling and warming: it is reciprocal to the Cv and Cdv so they move to the left into the area of the lower concentrations. Thus, there are 2 basic and reciprocal ways of achieving VF: i) by raising the concentration, and as result, the viscosity of the intra- and extracellular milieu at relatively moderate and even slow rates of cooling but very high concentrations VFAs, 20 Current Frontiers in Cryobiology which is defined as equilibrium VF and ii) by increasing the rate of cooling with a few or not at all exogenous VFAs present so deleterious intracellular ice formation is not achieved due to lack of time for growing ice crystal nuclei (kinetic VF).

That is how the concept of the Universal Cryopreservation Protocol was born (published first in the “Embryomail” in the spring of 2010). Kinetic Vitrification of Spermatozoa of Vertebrates: What Can We Learn from Nature? 04 40% 20% 0% Human Mouse Rat Fig. 7. 25 M sucrose (red) directly into LN2. Two methods of cryopreservation are compared. 28 Current Frontiers in Cryobiology Fig. 8A. An attempt to vitrify sperm of the polar bear, from left to right: sperm retrieval; fresh; slow frozen; and vitrified sperm.

2008). " Reproduction 136(2): 167-73. , Katkov I. I. &Kreienberg R. (2010). Cryopreservation of spermatozoa: Old routine and new perspectives. In: Principles and Practice of Fertility Preservation. Eds:J. Donnez and S. S. Kim. Cambridge, UK,, Cambridge University Press: Ch. 14, pp 177-99. , Risopatron J. &Sanchez R. (2012). Vitrification Technique- New Possibilities for Male Gamete Low-Temeprature Storage. In: Cryopreservation / Book 1 (ISBN 979-953-307-300-1). Eds:I. I. Katkov, inTech. 1: Chapter 2 (in press).

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