Untersuchungen zur Kryokonservierung humaner Adipozyten zur mehrzeitigen autologen Transplantation
The autologous transplantation of cryopreserved human adipocytes is a usual technique in Plastic Surgery, up to now. There are no evidences, that these cells have normal vitality and a complete integrity of their membranes, at the moment of the transplantation. Aim of this work was, to make a quantitative statement about the cell-vitality and cell-integrity of mature human adipocytes, by using an enzyme-based test-system. Therefore, the existing MTT-, XTT- and GPDH-tests were modified for the use of human adipocytes. For the protection of human cells, during cryopreservation, cryoprotective agents are a common group of chemical substances, in science. In this work we used hydroxyethylstarch, glycerol and a mixture of glycerol, dextran and polyvinylpyrrolidone. The vitality of the cells was measured before the cryopreservation, at -20°C and -80°C, and after 2, 7 and 14 days as well as after 1, 6 and 12 month. As an intermediate-result we ascertained, that a cryopreservation of human adipocytes for 48 hours, without using a cryoprotective agent, leads to a vitality-loss of 94,9% (MTT-test, -20°C) and after one year of cryostorage to a vitality-loss of 97,4% (MTT-test, -20°C). Comparable results were detected in the XTT- and GPDH-test as well as by using -80°C as cryopreservation temperature. The use of the selected cryoprotectives was able to reduce the loss of cell-vitality and the cell-membrane damage, which was measured by using the GPDH-test. The best quantitative result was achieved by using the cryoprotective mixture. After one year of cryopreservation the cell-vitality was increased up to 30% in the MTT-test and up to 60% in the XTT-test; the GPDH-test could prove a reduction of the loss of cell-integrity up to 50%.1. The inset of cryopreserved human adipocytes, without using a cryoprotective agent, should be no subject of practical use in medicine.2. Through the addition of cryoprotective agents, it is possible to improve the vitality and integrity of human adipocytes high significantly, in comparison with the non-use.3. The suitable cryoprotective was a mixture of one intracellular (Glycerol) and two extracellular (Dextran and Polyvinylpyrrolidone) protectives.
info:eu-repo/classification/ddc/610, Autogene Transplantation, Fettgewebe, Kryokonservierung, adipocyte viability, autologous fat transfer, cryopreservation, cryoprotective agents, enzymatic measurements of adipocyte viability, Adipozyten, Kryoprotektive, Medizin
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