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ИСТИНА ФИЦ ПХФ и МХ РАН |
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Successful application of human pluripotent embryonic stem cells (hESCs) in different fields of molecular and cell biology, as well as solutions for medical application tasks, unable without preparing of a large numbers of such cell material. Cryopreservation is a key operation in this process, because it allows to store a large amounts of cells for a long time and facilitates a process of their transportation. The standard protocol for cell cryopreservation is slow cooling. It is quite simple, but hESCs shows a poor survivability after thawing. The low survivability of hESCs after slow cooling is a result of different damaging effects such as the impact of dissociation solutions and components of freezing media, or because of wrong choice of freezing conditions which leads to damaging of cell membrane by ice crystals. Dimethyl sulfoxide (DMSO) is one of the most commonly used solvents in drug screening and it also used as a cryoprotective agent for cryopreservation of hESCs. So the influence of short-term exposure of DMSO on hESCs was analyzed. It has been shown that incubation of cells at 370C for 10 min in 10 % DMSO induces the cytoskeleton rearrangements, detaching from the substrate and dissociation of hESCs, while feeder fibroblasts remain on the culture surface and shows weak cytoskeleton rearrangements.