Respiration and glycolysis in the human diploid cell strain WI-38

Assessment of the respiratory and glycolytic capacity of non-growing WI-38 cells shows that, in the absence and presence of added glucose, the mean rates of oxygen consumption were 247 (QO₂ = 5.61) and 208 (QO₂ = 4.73) mμmoles/mg dry wt/hr., respectively. Mean glucose consumption was 225 mμmoles/mg dry wt/ hr. With uniformly labeled ¹⁴C glucose as substrate, 36 mμg atoms of carbon dioxide were produced, corresponding to 15–20% of the total cellular respiration. Mean values for lactate production in the presence and absence of glucose were 345 (Q_L^O2 = 7.85) and 196 (Q_L^O2 = 4.45) mμmoles/mg dry wt/hr., respectively. Human diploid cells in culture age, in the sense that their ability to proliferate decreases with time during serial subcultivation. Studies of their respiratory and glycolytic capacity as a function of the aging process showed that total respiration, glucose respiration and glycolytic capacity were relatively constant for cells in the middle and late passages and indicate that aging in this sense is not directly related to the respiratory and glycolytic capacity of the cell.     

Immunochemical quantitation of enzymes in human diploid cell line WI-38

A quantitative immunochemical study was carried out of four enzymes, cathodal esterase, acid phosphatase, glucosaminidase and β-glucuronidase. In homogenates of the human diploid cell line WI-38, the relative amounts of the enzymes increased with the passage number of the culture, although great variation was found in later passages just before death of the culture.     

Growth characteristics of bovine herpesvirus 1 (infectious bovine rhinotracheitis) in human diploid cell strain WI-38.

IBR virus was found to replicate in WI-38 cells. At a high input multiplicity the virus yield was comparable to that obtained in bovine cells, but comparable degree of CPE took longer to achieve. At a low input multiplicity of IBR virus, such as may be encountered in virus contaminated bovine serum, virus yield was only about 1% of that in bovine cells, with 50% of the cells showing CPE, followed by cell regrowth. Infectious virus was not recoverable from the regrown cells by 5 weeks after initial infection, and these regrown cells were susceptible to reinfection with IBR virus. Aging of WI-38 cells in cultures for as little as 1 week reduced IBR virus yield to 90% less than the yield from the same lot of cells inoculated 7 days earlier.     

A study of the cytogenetics of the human cell strain WI-38

Summary Cultures of the human diploid cell strain WI-38 were subcultivated under conditions which would meet the requirements proposed for the use of this strain as a substrate for the preparation of viral vaccines and would be in keeping with efficient production procedures. For chromosomal analysis, the cultures were combined in three groups at low, intermediate, and high passage levels, the latter being beyond those recommended for vaccine production. At all passage numbers, the incidence of aneuploid cells was low and constant up to those passages where the finite life span was approached and the population doubling time became markedly prolonged. At all passage levels, the incidence of gaps was higher than that of breaks but there was no significant increase of either of these abnormalities with continuous subcultivation. Among structural abnormalities dicentrics, despiralizations and deletions predominated. A significant increase in polyploidy occurred in the highest passage numbers, although the ratio of polyploidy to endoreduplication was constant throughout the series. Neither heteroploid transformation nor nonrandom chromosomal aberrations were observed. Nor was there correlation between observed aberrations and their location on the chromosomes. The incidence of hypodiploidy was lower than reported by other investigators. At the cellular level, no morphological changes could be associated with the distribution of chromosomal aberrations. This study was assisted by funds provided by Canadian Public Health Research Grant 605-7-710 of the National Health Grants Programme.