X-Message-Number: 33172 Date: Sat, 1 Jan 2011 10:55:10 -0800 Subject: Re: Cryopreservation since 1990 From: Brian Wowk <> Mike expressed the opinion that the quality of cryopreservations have gone down since 1990 despite the use of better cryoprotectant solutions because a lower percentage of people are being perfused with the solutions. Alcor did 10 cases in 2010. One was a post-mortem signup, one had cryopreservation blocked by relatives, and one was a middle aged man who suffered sudden cardiac death. The last two cases required court action to gain access to remains. The remaining seven, Alcor's last seven cases, all received cryoprotective perfusion. Six of those received standbys. That is as good as the Darwin/Leaf record of 1980s, and at a much higher case density. Three of these seven cases occurred within a one month period. Mike expressed concern that ice blockers in modern vitrification solution could worsen freezing injury in poorly-perfused areas by delaying ice nulceation to very low temperatures where water would be less mobile, and membranes less permeable to water. However the high molarity glycerol solutions used in cryonics in the 1990s were already doing this. The 7.4 molar glycerol solution in Mike's 1995 canine study that showed excellent preservation would have had a melting point near -50 degC, and would have likely supercooled tens of degrees below that before starting to freeze as highly concentrated cryoprotectant solutions tend to do. An attempt was made at 21CM to study what would happen to a brain cryopreserved and rewarmed after perfusion with only 80% normal concentration of M22. The experiment failed to elucidate what happens to tissue that freezes due to perfusion with sub-vitrifiable concentrations of M22 because even at only 80% normal concentration, no freezing injury was observed. It may be that the concentration regime in which freezing differences with and without ice blockers become most apparent is at dilute concentrations, well below full concentration; concentrations at which ice nulceation would normally occur near 0 degC, but becomes delayed to low sub-zero temperatures by ice blockers. The concern is valid in theory, but there is no data, and as far as the brain is concerned it would seem to only apply to a minority of patients when perfusion goes poorly. I remember speculating years ago that if a perfusion were going poorly it might be theoretically beneficial to add ice nucleators to the perfusion solution. However it would be a difficult call to make, and there is no data to support to it. ---BW Rate This Message: http://www.cryonet.org/cgi-bin/rate.cgi?msg=33172