**8***.* **Conclusion:** *"Race for the Pace"***: Is the universal cryo-protocol possible?**

The universal cryoprotocol, that would fit *all* types of cells, at least if they are in suspension on make a thin layer, would be the *Holy Grail* of cryobiology. Here is our hypothesis for consideration [Katkov, 2010]:


Kinetic Vitrification of Spermatozoa of Vertebrates: What Can We Learn from Nature? 31

*"Here we can only note the irony that, having been launched by breaking free of Luyet's tyranny of ultrarapid cooling, vitrification methods have now essentially turned back closer to Luyet's original idea of cooling as quickly as possible with minimal intracellular exposure to cryoprotectants* [i.e., kinetic vitrifcation VF, *I.I. K.*]*,*, *albeit this time using at least marginally adequate concentrations of intracellular solutes. The ghost of Luyet lives on in the form of this ongoing methodological evolution, and we think he would have been pleased to see how his ideas about vitrification ultimately related to the now widespread use of vitrification as a practical and successful method of cryopreservation long after he, himself, had abandoned this approach."*[Fahy & Rall, 2007] As a cryobiologist, who has been working in the same field, the Editor (at the Eves of 2012)


Had that all happened, Father Luyet would have been indeed thrilled to see how his

And for me, it is better to live under "tyranny" of the genius than under "ochlocracy" of the ignorant or "democracy" of the arrogant, and seeing as Wikipedia and other internet resources have been invaded (and infested) by cryonics "experts", and are full of their biographies, cryonic companies' descriptions, etc, while a reader can find neither biographies of Father Lyuet nor other prominent cryobiologists, both who passed away and live and in good health today. Unfortunately, Wikipedia has failed to

But that would be a topic of our other story, here we must stop and say just only that: kinetic vitrification of sperm, the early child of Father Luyett and the other *pioneers of the cryobiological frontiers*, is very much alive and on the march! And our own success and failures, honestly described in this Chapter, have only strengthened the position K-VF


might announce the following "resolutions":

of his science), but his body as well;

**papers, which are not cited there** 

equation (25) in our paper [Katkov & Levine, 2004].

 

is replaced by the VTF equation in [Fahy & Rall, 2007] as following:



method has been spread and are opening new horizons!

be fair and balanced on this matter, but hope it'll change with time.

as a viable (not marginal!) and very promising method of cryopreservation.

**Appendix 1. Some peculiar similarities between [Fahy & Rall, 2007] and our earlier** 

We invite the readers to compare the physical description of vitrification in that chapter by Fahy and Rall, particularly sub-chapter *"The kinetic basis of vitrification"*, and the first part of the *"Optimal storage below Tg"* with our preceding publications (pp. 71 and 75 in [Katkov & Levine, 2004] and pp. 353-4 "*6. Storage at temperatures higher than Tg of water"* in [Katkov *et al.*, 2006] respectively). The only substantial difference is that "*the most widely used"* WLF

( ) ( ) ( ) ( )10 ( )

() ( ) *<sup>v</sup>*

 *T Te <sup>g</sup>*

*T Tg g*

1 1 2 2

> *B T T*

 *T e* 

( ) 2.303 ( )

*g g g g C TT C TT C TT C TT*

(1)

, (2)


Thermodynamic analysis of the most recent attempts of creating novel systems for kinetic VF such as cryogenic oscillating heat pipes [Jiao et al., 2006; Jiao et al., 2009], nano-droplets [Demirci & Montsesano, 2007], quartz capillaries [Risco et al., 2007], and some others approaches that claim *"ultra-fast"* rates (see a comprehensive review by Criado in this Book), which in our opinion, do not produce the rates fast enough to reach the majority of B*cr*'s without using exogenous permeable (and thus, potentially toxic) vitrificants. Thermodynamical considerations that prove this statement are not in the scope of this Chapter and will be done elsewhere. In fact, the *hyper*-fast rates of cooling and warming will be needed, and there is about of an order or two of magnitude difference between *"ultra-"* and *"hyper-"* (cf. *ultrasonic* and *supersonic* speed of flight as an example).

Introduction of such a *"Universal Kinetic Vitrification Protocol*" applicable for *all* cells (at least for those that are in suspension or make a thin attached layer) would shift the whole paradigm in cryopreservation of germplasm (and other types of suspendial cells) and in cryobiology as a science. It will require both new equipment for realization of hyper-fast rates (on which we are working now) and new methods of measurements. For example, it is not clear how *Th*, *Tm*, and *Td* curves on the Fahy diagram would behave at speeds of cooling and warming in order of thousands OC/min, and how that could be measured: they may disappear completely! In any case, it will open not only the possibility of development of a uniform protocol and equipment for all existing and (which especially important) *new* types of cells and species, but it will also bring new, very challenging but exciting horizons for basic cryobiology as well.
