Preservation of Avian Cells at Sub-zero Temperatures

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Abstract
The cryoprotective agents dimethyl sulfoxide (DMSO), glycerol, polyvinylpyrrolidone (PVP) and dextran were evaluated for their ability to protect avian cells during storage at sub-zero temperatures. DMSO was the most effective cryoprotective agent for the short- and long-term storage of avian cells and glycerol was also effective when used at low concentrations. PVP and dextran did not protect avian cells during storage. Primary chicken cells and avian cells at higher passage levels were successfully recovered after storage with DMSO for periods ranging from 4-12 mo. Whole embryos were frozen with varying concentrations of DMSO, and the survival rates and cell yields were determined for chickem embryo fibroblasts (CEF) prepared from the frozen embryos. Frozen embryos did not yield as large a number of cells as the fresh ones, but storage with 25% DMSO gave over 50% cell yields and survival rates. The ability of frozen CEF and chicken embryo kidney cells (CEK) to support virus growth was also investigated. Assays of infectious laryngotracheitis, Sindbis, fowlpox, Newcastle disease, adeno- and turkey herpes viruses agreed within 0.5 log10 (50% tissue culture infective doses) when conducted in frozen and fresh CEF and CEK.