10.1007/s11827-008-1004-9

Abstract

Structural and functional changes in cytoplasmic membranes and cell organelles play a crucial role in cell damage at low temperatures. These changes are reversible if adequate measures are taken to protect biological membranes against cold-induced injuries. In this study, the possibility of cryopreservation of membrane integrity by long-term storage of samples at low temperatures (−80°C) is demonstrated using differentiated nuclear blood cells as an example. In addition to classical methods, freezing of human leukocyte suspensions was carried out in a novel nontoxic cryoprotecting solution under a newly developed exponential freezing program based on the use of cryoprotectors and repair additives. This program ensures the same cryopreservation effect as the linear program, but is economically more efficient and less labor-consuming. After exponential freezing in a cryoprotecting solution and storage at −80°C, blood leukocytes retain their eosin resistance (91 ± 5% of thawed leukocytes) and phagocytic activity (76.7 ± 14.7% of thawed neutrophils) for 180 days. The novel technology of cell cryopreservation employing a nontoxic cryoprotecting solution, exponential freezing program, and fast thawing is economically efficient, easy to perform and applicable to storage of any animal cells.