CREATE ACCOUNT

Request Registration Code

Terms of Service

All messages posted at this site express the views of the author, and do not necessarily reflect the views of the owners and administrators of this site.

By registering at this site you agree not to post any messages that are obscene, vulgar, slanderous, hateful, threatening, or that violate any laws. We will permanently ban all users who do so.

We reserve the right to remove, edit, or move any messages for any reason.

  I agree to the terms of service

FORGOT YOUR DETAILS?

CI Report: Pichugin Research Findings Lead to New Washout Solutions

by Robert C.W. Ettinger
January 2002

As a result of some of the research of Dr.Yuri Pichugin, the Cryonics Institute's Director of Research, changes have been made in the washout solution and the perfusate vehicle solution. Following are the compositions of the new solutions, which yield significantly better results than the previous solutions.

In addition, there had been discussion, in-house and with various consultants, over the merits of solutions or ingredients used or recommended by others. In particular some had recommended use of HES instead of mannitol in the washout solution, and some had recommended use of organ-preservation solutions such as Viaspan and its successors and competitors, including a solution known as m-RPS-2, a modified renal preservation solution, which differs from the new CI vehicle solution in that it contains glutathione. Direct comparisons were made, as summarized below, and the suggested alternatives were found not to be better than the new solutions now adopted.

The following is my summary, and not Dr. Pichugin's full report, but I think I have avoided errors and included the essentials.

A. The new CI Washout Solution:

 

mM/L

Molecular Weight

grams/L

KCI

30.0

74.55

2.24

NaHCO3

10.0

84.01

0.24

KH2PO4

7.2

136.1

0.98

MgCl2•6H2O

2.0

246.48

0.49

Glucose

180.0

180.20

32.44

Mannitol

50.0

182.17

9.11

 

To adjust pH of the solution to 7.6,dissolve1g KOH in 2 ml of distilled water and add 15 drops of this to each liter of solution.

B. The new CI perfusate vehicle solution (to which glycerol is added in phased steps):

Same as above except that there is no mannitol.

C. Comparison of new CI washout solution with one using HES (50 g/L) instead of mannitol:

Rat brain hippocampal slices were incubated at 29ºC for one hour before use. Slices were then kept in the CI or comparison solution for 4 hours at 0ºC, then incubated in the Oslo chamber for one hour for recovery, then tested for K/Na ratio in the flame spectrometer.

Control slices showed an average ratio of about 2.83, with standard deviation of about 0.28 in the first series. After being in the CI washout solution as noted, the ratio in the first series averaged 2.36, S.D. 0.15 -- viability slightly less than controls. In the solution with HES the average ratio was 2.55, S.D. 0.22. There was no statistically significant difference between the solution with mannitol and the solution with HES. A repeat series of each yielded essentially identical results.

Comparing the new CI vehicle solution with m-RPS-2 was done similarly, with nearly identical results: no significant difference.


TOP