В данной работе было исследовано содержание трансформирующего фактора роста TGF-β1 и нейротрофического фактора мозга BDNF в зоне лимба роговицы после введения криоконсервированных ядросодержащих клеток кордовой крови (кЯКККЧ) в экспериментальной модели лимбальной недостаточности роговицы (ЛНР). Криоконсервирование ЯКККЧ проводили с высоким или с существенно сниженным содержанием декстрана. Было показано, что независимо от метода замораживания при введении кЯКККЧ животным с индукцией ЛНР в ткани роговицы наблюдается достаточно высокое содержание BDNF и TGF-1b, что способствует улучшению регенерации клеток лимба, эпителия и стромы роговицы глаза.
Cryopreservation of cord blood nucleated cells for subsequent application in clinical practice is the only way of their long-term storage. The aim of this study was evaluation of the apoptosis stages and phosphatidylserine distribution in the nucleated cells membrane after cryopreservation by different methods. Nucleated cell fractions were frozen under the protection of the cryoprotectants with different mechanism of action. It was shown that nucleated cells isolation in polyglucin and subsequent freezing under 5% DMSO protection, as well as cells isolation by two-step centrifugation method and subsequent freezing under 10% of polyethyleneoxide (PEO) protection, allowed to keep intact most of the cord and peripheral blood cells. Cryopreservation of nucleated cells isolated using Ficoll, regardless of the used cryoprotectant, leads to significant disruption of the lipids asymmetric distribution in membrane and significantly reduces the number of living cells. It has been found that cord blood nucleated cells more resistant to damaging factors of cryopreservation than peripheral blood cells, that is shown in significant differences between the number of live cells in quite all cases before and after cryopreservation.
New efficient methods for separation and cryopreservation of nucleated cells from umbilical cord blood, which allow you to save a quantitative and qualitative composition of nucleated cells (including hematopoietic stem] after discharge, and demonstrate a high level of preservation of structural and functional properties and viability of CD45+ and CD34+-cells after cryopreservation.
