DEVICE AND METHODS FOR THE CULTURING, STORAGE AND CRYOPRESERVATION OF STEM AND DIFFERENTIATED CELLS OF HUMAN AND ANIMAL ORIGIN

摘要

In order to improve preservation of the structure and functions of human and animal stem cells with varying levels of differentiation and of highly differentiated tissue cells during in vitro and ex vivo culturing and during cryopreservation and long-term storage under standard and extreme conditions, culturing media are used that correspond to the cell type in question and to which one or more gels of varying density are added that contain a submicron emulsion of chemically and metabolically inert perfluoroorganic compounds (PFOC). PFOC gel structures perform the function of a skeleton and at the same time imitate the intercellular matrix and thus may serve as a substrate and gas exchange medium. The gels are composed of emulsions of individual PFOCs or emulsions of mixtures of PFOCs, the dispersion of which is stabilised by non-ionising surfactants (NIS) based on block copolymers of polyethylene oxide and polypropylene oxide and derivatives thereof of different molecular weights and component ratios. Depending upon their density, gels containing submicron emulsions of PFOCs may advantageously be used for the culturing, storage or cryopreservation of cells according to their density. Gel density may be reduced by introducing a saline solution medium containing low-molecular weight osmolytes, and conversely the density of the gel may be increased by increasing the concentration of basic components, i.e., PFOCs and NISs, by partial lyophilisation and/or by centrifuging. Cell cultures in suspension and in monolayer growth following cryopreservation, storage or culturing in PFOC-containing gels have a viable cell average of 90% and are easily removed from the substrate gels, sub-culture under standard conditions, divide, and preserve full functionality and their differentiating capabilities.