Effects of Physiological Isotonic Cryoprotectants on Living Cells During the Freezing-Thawing Process and Effects of Their Uptake by Electroporation: Sp2 Cells in Alginate-Trehalose Solutions

摘要

In the present study, effects of isotonic alginate and trehalose solutions on electroporated and unpulsed control murine myeloma cells (Sp2 line) during the freezing-thawing process were evaluated. To investigate the effects of these potential cryoprotectants, microscopic examination of cell suspensions in alginate-NaCI, algi-nate-trehalose, or trehalose media was performed during freezing-thawing at various cooling/warming rates (e.g., -1, -5, and -90 ℃/min). It was found that, even at the lowest cooling rate, extracellular ice grains did not mechanically suppress cells in the trehalose-substituted medium. Moreover, at all cooling rates, intracellular ice nuclea-tion seldom occurred in the presence of extracellular trehalose.