| Abstrakt: |
In these freezing experiments utilizing glycerol and sucrose as additives, the primary criterion employed for evaluating maintenance of the Ehrlich-Lettre hyperdiploid carcinoma cells was oxygen consumption, endogenously and with added lactate or glucose. Microscopic examinations were made to determine the morphologic state of the cells, and viability determinations carried out with mice. Glycerol and sucrose were equally effective for preservation of the cells for a few days, but sucrose was more effective when cells were kept for many weeks. Optimum conditions in our experiments were: (a) adding sucrose to the washed cells to make a concentration of 0.4 M, (b) reacting with the cells for 10 min, (c) freezing at a rate requiring 180 sec to reach —79°, (d) maintaining the frozen cells at —79°, and (e) thawing the cells at +56° for 1 1/2 min. These cells, following a twofold washing with isotonic phosphate buffer, were found to have near to normal oxygen consumption, morphology and viability. |
