Perfected Brain Slice Cryopreservation now in Sight

From: Paul Wakfer (
Date: Sat Jul 08 2000 - 12:32:38 MDT

        At the recent Fourth Alcor Conference on Life Extension Technologies
Dr. Greg Fahy presented current experimental results from the
Hippocampal Slice Cryopreservation Project (HSCP) of which he is chief
scientist and co-principal investigator. Now in its second year of
operation, the HSCP is being conducted at a research institution
associated with a major Southern California university by Yuri Pichugin
as experimentalist, and is funded by the Institute for Neural
Cryobiology (INC). Since that presentation, confirmation of those
results and additional progress have been made.

        The following research milestones have now been reached:

1. Stable and reproducible experimental procedures have been perfected
for dissecting, handling & maintaining rat hippocampal slices with
minimal harm, for vitrification solution (VS) addition & removal, for
cooling, vitrification & rewarming, and for assaying cellular viability.
2. Using a potassium/sodium ratio assay for cellular viability (cells
need to actively maintain this ratio in order to stay alive), addition &
removal of the VS currently used in the HSCP yields viabilities in the
range of about 50% of the viability of fresh, untreated control slices.
Slices vitrified in this VS have shown no injury attributable to
vitrification & warming per se in two separate experiments, and the
reproducible cellular viability of such vitrified hippocampal slices is
now at 53% of untreated controls. The primary challenge, then, is to
reduce injury associated with the VS itself.
3. Work is about to begin with a new VS which has allowed 21st Century
Medicine researchers to achieve 95% cellular viability after testing the
VS on rabbit kidney slices. In addition, work is continuing on methods
of adding & removing VS that may be less damaging. Finally, it is
possible that longer recovery times may further improve the ultimate
viability of the slices. All in all, prospects for further improvement
appear excellent.

        The ultimate goal of the HSCP is perfected cryopreservation of brain
hippocampal slices. This requires not only high cellular viability, but
clear demonstration of functioning cellular interconnections. More
details about this advanced global viability assay may be found at the
INC website listed below. Once high cellular viability is achieved
(which now looks to be very realistic before the end of the year 2000),
work will begin on that final clear demonstration.

        The many generous donors of well over $100,000.00 spent so far to
support this project are to be commended for their judgment and
foresight. However, more money is now needed if the HSCP is to continue
its success and complete its original project goals, which now more than
ever before appear to be fully attainable.

        Please consider donating money to support the continuation of this
important work which in all likelihood will hasten the achievement of
perfected, fully-reversible, long-term human cryopreservation.

        INC is a registered California Public Benefit Corporation (501(c)(3) -
charitable) and all donors receive an official receipt which may be used
to reduce US and California taxable income.

        Checks or other financial instruments should be made payable to
Institute for Neural Cryobiology and mailed to the address below. Any
amount is welcome.

        The Institute for Neural Cryobiology
        238 Davenport Rd #240
        Toronto, ON M5R 1J6 Canada
        Phone:416-968-6291 Fax:559-663-5511

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