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KMID : 0380319700040000085
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Journal of Korean Research Institute for Better Living
1970 Volume.4 No. 0 p.85 ~ p.95
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WHEY MEDIUM FOR CULTIVATION OF CHICK EMEBRYO CELLS AND EFFECT OF WHEY ON JAPANESE ENCEPHALITIS VIRUS
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Abstract
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Animal serum nas been a necessary constiuent of most media for continuous cell cultures. However, many sera contain antibodies and other inhibitors of microorganisms. It would be desirable to replace serum with another substance which does not contain such inhibitors and which sustains cells in a condition sensitive to viral effects for long-term observation. Such a medium would be not only useful in the studies of nutritional factors, but also for comparative assay of a variety of viruses on a number of different cell cultures, for safety testing of virus vaccines, and for isolation of viral agents in tissue culture.
The present report concerns the nutritive value of whey and its effect on Japanese encephalitis (JE) virus chick embryo tissue culture media devoid of serum. Whev was prepared in various ways. Whey media included whey with and without gelatin,yeast extract, Tween 80, polyviny1 pyrrolidone(P.V.P), and vitamin B＿12 as supplements in Hank´s balanced solution or M199 plus 0.5% lactalbumin hydrolysate. Each tube or bottle was planted with 1.6×10^6 cells per 1.0 ml of the media. Growth was measured by the method of protein estimation with the Folin phenol reagent. The results of these experiments are as follows:
1. Whey heated at 100˚C for 5 and 10 minutes respectively stimulated the growth rate of chick embryo embryo cells markedly at 10% concentration in a basal medium. Unheated whey was more effective.
2. Whey(10%) treated with pepsin yielded results similar to those with heated whey.
3. The whey medium to which was added gelatin, yeast extract, Tween 80, p.v.p.and vitamin B＿12 respectively did not show any advantage over the original whey medium.
4. Chick embryo monolayer cells were maintained in whey media for periods of up to 18 days and in M199 containing 10 percent whey for periods of up to 40 days. with out replacing the fresh media and without marked PH drop.
5. Protective effect of whey for JE virus particles eas almost idential when compared with animal serum in PBS as a viral diluent at the room temperature.
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