Critical cooling and warming rates to avoid ice crystallization in small pieces of mammalian organs permeated with cryoprotective agents.

Measurements were made by differential scanning calorimetry on small pieces of rabbit kidney permeated with 2, 3-butanediol containing mainly the levo- and dextro-isomers. The critical cooling rates necessary to vitrify the pieces of organ, and the corresponding critical warming rates which are required to avoid crystallization in the vitrified samples, were determined. The dynamic method used for these determinations is described. The glass-forming tendency and the stability of the amorphous state were both greater in the kidney tissue samples than in the bulk cryoprotective solution. This result is discussed in the context of the lowering of the freezing point of water in emulsions and the promotion of supercooling in hydrogels and porous materials. In corresponding experiments with rat hearts impregnated with 1,2-propanediol, only the critical warming rate was reduced.

Effect of Saccharides on the Glass-Forming Tendency and Stability of Solutions of 2,3-Butanediol, 1,2-Propanediol, or 1,3-Butanediol in Water, Phosphate-Buffered Saline, Euro-Collins Solution, or Saint Thomas Cardioplegic Solution

The effect of sugars or reduced saccharides trehalose, sucrose, sorbitol, or mannitol on the glass-forming tendency during cooling and the stability of the wholly amorphous state during warming has been studied with 2,3-butanediol, 1,2-propanediol, or 1,3-butanediol in three different carrier solutions. The 2,3-butanediol contained 96.7% (w/w) racemic mixture of the levo and dextro isomers and 3.1% (w/w) of the meso isomer (called 2,3-butanediol 97% dl). The carrier solutions were water, a phosphate-buffered saline, and two organ preservation solutions (Euro-Collins and Saint Thomas). The latter two were chosen because they are often used for kidney and heart preservation, respectively. The concentrations of 2,3-butanediol, 1,2-propanediol, and 1,3-butanediol varied respectively from 25 to 34, 30 to 35, and 30% (w/w). The concentrations of saccharides were 4 or 5% (w/w). In the absence of saccharides, for a given 2,3-butanediol concentration, the glass-forming tendency increased in the following order: water, Saint Thomas, the phosphate buffer, Euro-Collins. Addition of 4 or 5% (w/w) saccharide resulted in a large increase in the glass-forming ability of the solution during cooling and increased the stability of the glass during warming; but replacement of 4 or 5% diol by an equivalent weight (percentage) of a saccharide decreased, though to a lesser extent, these properties.

[Organ cryopreservation: current status of research carried out in Grenoble]. / Cryopréservation d'organes: etat actuel des recherches développées à Grenoble.

After discussing the problem of organ cryopreservation and reviewing the current data available on this subject, the Grenoble project is presented. Physical and biological studies have been combined with experimentation of autologous renal transplantation in rabbits to assess the functional value of the retransplanted organ after treatment and cooling. Renal resistances are measured during perfusion of the kidney with the cryoprotective solution. In order to verify the homogeneity of the cryoprotector concentration in the organ, on NMR spectral imaging test has been developed. A new rapid imaging method now allows real time monitoring of concentration variations during perfusion. In addition to concentration and homogeneity, analysis of local spectra also provides information about the local temperature de the kidney.