Cultivation of human epithelial cells in tissue culture.

Abstract

1. Experience is described in the serial subcultivation of 2 human cell lines derived from conjunctiva and kidney and established in serial passage in vitro by Chang. 2. Thirty-eight human sera were compared in their ability to support multiplication of the 2 cell lines. Two of the sera were toxic to both cell lines. The remaining 36 sera were non-toxic but varied in their ability to support cellular multiplication. Horse and calf sera were superior to rabbit, sheep, and chicken sera but inferior to the human serum control. 3. Comparison was made of the multiplication of the 2 cell lines when cultivated in Chang's, Eagle's, and 199 media with added human, horse, or calf serum in 10 and 20% concentration. 4. Conjunctiva cells have been serially subcultivated for 18 passages in media containing horse or calf serum but no human serum. Kidney cells have been carried for 14 passages in such media. 5. Conjunctiva and kidney cells survived for varying periods when stored at 32°C or 5°C. Some sedimented conjunctiva cells survived 10 freezing-thawing cycles and formed a sheet of cells when incubated at 37°C. 6. Three of 11 conjunctiva cultures survived lyophilization and revived when re-incubated at 37°C for 5 weeks. No contaminating bacterium or virus could be detected in these cell lines by the injection of nonviable cell extracts into tissue cultures of monkey kidney, embryonated eggs, rabbits, guinea pigs, and mice. 7. A method is described for the treatment of contaminated tissue cultures. 8. The conjunctiva cells gave results comparable to monkey kidney cells when used for isolation of poliomyelitis virus from 2 stool specimens.