R. A. Bonar

The behavior of HeLa-S3 cells under the influence of supranormal temperatures

HeLa cells exposed to environmental temperatures higher than the usual 37°C undergo marked morphological and biochemical changes which are reversible after return to normal temperatures. Exposures to 41°C had little effect, but at 42 to 43°C changes were evident by 30 minutes and pronounced at 1–2 hours. At 44–45°C degenerative changes and cell death were frequent. At 42–43°C cell response was characterized by partial loss of the granular component of the nucleolus with retention of its fibrils. Synthesis of the 45 S ribosome precursor RNA was partially inhibited at 43°C as was the conversion to ribosome RNA of precursor molecules formed at 37° and then metabolized at 43°. The number of perichromatin granules was markedly increased in the nuclei of heated cells and threadlike structures of the approximate diameter and density of the peri-chromatin granules appeared. Mitochondria and Golgi apparatus remained essentially unchanged morphologically but the polysome arrangement of the ribosomes was completely lost and they were distributed as monosomes after heating. Cell recovery from exposure to 2 hours at 43°C was complete within 24 hours, the only alteration remaining being an increase in the number of lysosomes.

Ultrastructure of viruses of myeloblastosis and erythroblastosis isolated from plasma of leukemic chickens.

Summary Ultrathin sections of the pellets formed by ultracentrifugation of plasmas from birds with myeloblastosis and erythroblastosis have been examined by electron microscopy. In the material from both diseases there were observed structures of similar appearance represented by images of round or oval shape consisting centrally of a dense core of about 40 mμ diameter. These were surrounded by a less dense material and were limited by an outer membrane-like structure with a total particle diameter of about 80 mμ. In many instances the limiting membrane appeared to be double. The ultrastructure of these agents of the avian leukemic diseases was very similar, if not identical, to that of the viruses of other chicken neoplasms. No particles of these characteristics were obtained from the plasmas of apparently healthy chickens.

Lipids of the BAI strain A avian tumor virus and of the myeloblast host cell

Abstract Lipids of the BAI strain A virus and of the myeloblast host cell and nuclear, microsomal and external membrane cell fractions were analyzed by thin-layer chromatography. Total virus lipids consisted of 34% cholesterol, 5.3% neutral fat and 61% phospholipid. Of the latter, the principal components were sphingomyelin, 16; phosphatidyl ethanolamine, 21; and lecithin, 11%, respectively. This constitution of the BAI strain A virus was proportionally similar to that of the influenza agent. Virus lipids were qualitatively similar to those of the whole host cell and the nuclear, microsomal, and cell membrane fractions. Marked quantitative differences were observed between the virus and host cell components, particularly with respect to the cell membrane fraction in which the low content of cholesterol, 9%, and 30% neutral fat were the reverse of the virus with 34 and 5.3% of these constituents. Major differences observed also with the phospholipids were, again, greater in the comparison between the virus and the cell membrane. The findings were interpreted to indicate the improbability of major cell membrane contribution to the virus lipids. Consideration of the analytical data and correlation with the ultrastructural aspects of BAI strain A budding from the myeloblast surface suggest that the virus lipids are specifically assembled in the process of virus synthesis not only from the external cell membrane but also principally from contiguous and possibly other cytoplasmic structures.

Canine urinary bladder epithelial cells: Preparation for cell culture by enzyme dispersion

SummaryIn a qualitative and quantitative study of enzymic dispersion of cells from the mucosal layer stripped from canine urinary bladder, trypsin was found to be equal or superior to the other enzymes tested for dispersal of urothelial cells specifically. Collagenase or collagenase plus trypsin served to disperse the whole tissue. A procedure for recovering the urothelial cells as a single-cell suspension and establishing them in culture is presented. The morphology, culture behaviour, and chromosome complement of these cells is described.

In Vitro Chemotherapeutic Testing of Urologic Tumors

We studied 20 transitional cell tumors of the bladder and 25 adenocarcinomas of the kidney in vitro to determine their chemotherapeutic sensitivity. The different sensitivity patterns among the individual tumors were demonstrated. Identical drug sensitivity patterns could be identified in the primary and metastatic sites, and in tumor tissue removed from the primary and metastatic deposits in the same patient. Human renal adenocarcinoma maintained in the athymic mouse demonstrated identical chemotherapeutic sensitivity patterns in vitro and in vivo. Our data would support that these in vitro chemotherapy studies may assist in the selection of agents to use in human tumor-bearing hosts.

In vitro culture of epithelial cells derived from urogenital tissues

SummaryA tissue culture procedure is described which yields a high percentage of successful longterm cultures of epithelial cells derived from malignant and non-malignant human urogenital tissues.

BAI strain A avian (myeloblastosis) leukosis virus from myeloblast tissue culture.

Summary A combined culture-dialysis chamber was devised for producing BAI strain A virus in vitro in amounts greatly increasing the scope of physical and chemical studies of the agent. The yield of virus was approximately 15 mg dry wt/day/chamber. Infectivity of the culture virus was the same on a particle basis as that of virus from leukemic chicks. Properties of the culture virus were indicated by gradient centrifugation, gel electrophoresis and by the character of RNA extracted from the agent.

Establishment and characterization of a cell line (SS78) from a human renal cell carcinoma

SummaryA new cell line, SS78, was established from a primary renal cell carcinoma of a Caucasian male. The tissue was dispersed with collagenase, and viable cells were separated by flotation on a Ficoll-Hypaque gradient. In culture, the SS78 cells retained a distinct epithelial morphology, and no fibroblastlike cells were seen. The cultured cells were aneuploid with a modal chromosome number of 80 and had several marker chromosomes. Inoculation of the cultured cells into athymic nude mice caused tumors at the sites of inoculation.

Polyploidy in mammalian urothelial cells

SummaryThe mitotic indices and the extent of polyploidy in urothelial cells of baboons, dogs and swine were studied. All three species had very low mitotic activity in vivo but short-term culturing of these cells in vitro stimulated mitosis thus enabling chromsome counts. Tetraploid cells were found in the urothelium of all three species, and higher ploidies also in dog and swine. There were substantial differences in the proportions of diploidy and higher ploidies among the three species and among individuals within cells were diploid (68%). Baboon urothelial cells had only two ploidy classes and 92% were diploid.

Identification of Avian Erythroblastosis Virus by Precipitation with Chicken Immune Serum.

Summary Spheroidal particles of about 100-120 mμ diameter occur characteristically in the plasma of chickens with erythroblas-tosis. With formalin-inactivated concentrates of these particles, immune serums have been produced in the chicken which effectively neutralize the infectivity of the agent. These antiserums also cause agglomeration of particles in concentrates identifiable by electron micrography as those found characteristically in this disease. The results have been interpreted to indicate that the characteristic particles agglomerated constitute the viral agent of avian erythroblastosis.