Abstract
The cryoprotection of the sheep erythrocyte intermediate EAC4 cells, used as reagent in titration of the first complement component, Gl, was investigated. The cryoprotective agents tested were untreated polyvinylpyrrolidone (PVP), purified PVP, neutralized PVP and a hydroxyethylated potato starch of high viscosity, Avelex 1030, hydrolyzed for 40 min. Recovery of EAG4 cells after thawing was 80–90 %, with best results using untreated, purified or neutralized PVP. The EAG4 cells frozen in the presence of untreated PVP showed, however, increased susceptibility to the hemolytic action of Gl, whereas cells frozen with purified or neutralized PVP or with Avelex 1030 gave titers similar to that obtained with fresh cells. Gl titrations with frozen and thawed EAG4 cells gave more reproducible results than those obtained when titrations were performed with fresh separately prepared cells.
Keywords: sheep erythrocyte intermediate; EAG4 cells; freezing at –196°G; storing at –90°G; titrations of the first complement component, G1
Sammendrag
Bevaring af EAC4 celler som reagens for titrering av første komplementkomponent, C1, ble undersøkt etter nedfrysing til –196°C. Som kryoprotektive midler ble brukt ubehandlet polyvinylpyrrolidone (PVP), renset PVP, nøytralisert PVP og en hydroksyetylert stivelse (Avelex 1030) hydrolysert i 40 min. Cellegjenvinningen var uavhengig av hvilken PVP som ble brukt, mens Avelex 1030 ga litt dårligere resultater enn PVP. Den hemolytiske virkning af C1 økte ved bruk av EAC4 celler som hadde vært frosset i nærvaer av ubehandlet PVP, mens celler som hadde vsert frosset med renset eller nøytralisert PVP eller med Avelex 1030 ga tilnaermet samme titre i C1 titreringer som de ufrosne cellene. Gl titreringer med frosne EAC4 celler ga mer konstante resultater enn når forskjellige prepareringer av ufrosne celler ble brukt som reagens.
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