Thomas A. McCoy

Amino Acid Requirements of the Novikoff Hepatoma in vitro.

Summary Novikoff hepatoma requires 12 amino acids and glutamine for growth in vitro. When both glycine and serine were deleted from the medium, no growth occurred. In presence of serine some growth was apparent, but glycine was more effective in stimulating tumor cell proliferation. Glutamic acid and sodium thioglycolate could partially spare glutamine and cysteine requirements, respectively, and several sulfur-containing inorganic salts could partially replace cysteine. Arginine could be replaced by citrulline but ornithine was not active. When arginine was deleted, the typically rounded cell of Novikoff hepatoma was no longer apparent; but a predominance of fibroblasts remained attached to the surface of the flasks.

Growth-Promoting Properties of Pyruvate, Oxalacetate, and α-Ketoglutarate For Isolated Walker Carcinosarcoma 256 Cells

Summary When Walker carcinosarcoma 256 cells were grown under clonal growth technics. 50-100% plating efficiency was noted in a medium containing whole bovine serum. When dialyzed serum was used in the medium, few or no colonies were formed. Addition of pyruvate, α-ketoglutarate, and/or oxalacetate to the substrate resulted in plating efficiencies of 90-100%. Activity of these 3 metabolites was the same irrespective of addition of individual compounds or combinations thereof. Other Krebs acids and metabolites tested failed to promote survival and growth of isolated Walker cells.

A Glass Helix Perfusion Chamber for Massive Growth of Cells in vitro.

Summary Freshly excised Jensen sarcoma cells have been cultivated in a helix perfusion chamber. Growth was luxuriant and masses of tissue were clearly visible to the naked eye. Microscopic examination revealed healthy cells with a high degree of mitotic activity. This system is readily applicable for massive growth of cells which cannot proliferate in free suspension.

Two Nutritional Variants of Cultured Jensen Sarcoma Cells.

Summary Two nutritional variants (JA-1, JA-2) have been developed from Jensen sarcoma cells cultured in vitro. Cells of the parental Jensen sarcoma required asparagine for growth, but those of the variant strains grew readily in the absence of exogenous asparagine. This variation in nutritional requirement has been retained in JA-1 and JA-2 for 23 and 16 subculture generations in vitro and in 3 and 18 in vivo transplant generations, respectively. Thus, this change was stable and heritable under the conditions described. Some growth and histological characteristics of these variants were described.

Production of beak and skeletal malformations of chick embryo by semicarbazide.

Summary Semicarbazide · HCl when injected into the embryonated White Leghorn egg at 4-6 days produced shortened and malformed lower beak and bent tarsometatarsal and tibiotarsal bones. The embryos were most sensitive to this injection at 6 days of incubation. Gross and histological effects were quite marked 48 hours after injection. Nicotinamide, pyridoxal, pyridoxine, biotin, and riboflavin administered simultaneously with SCH did not overcome the effects. Different effects on bone and soft tissues were produced with 1-phenylsemicarbazide. p-Hy-drazinobenzoic acid, but not benzoic hydra-zide, produced abnormalities very similar to those following SCH treatment. Only a small incidence of skeletal defect resulted from administration of thiosemicarbazide.

Inhibition of development of chick embryo liver by aminoguanidine.

Summary Aminoguanidine sulfate when injected into the yolk of a 4-day embryonated egg produced a severe inhibition of liver development of the chick embryo when the specimens were examined 10 days later. Apparently, the parenchymal cells were the sites of inhibition as the surviving tissue, in addition to being reduced in mass, was composed largely of connective tissue and cystic structures. Other organs and tissues of the specimens appeared to be relatively unaffected. The most effective sub-lethal dose at 4 days incubation was ineffective when administered at 9-10 days of incubation. Other hydrazides possess peculiar effects which bear no resemblance to that of AGS.

Phosphomannose Isomerase Activity in a Spectrum of Normal and Malignant Rat Tissues.

Summary Homogenates of normal and malignant tissues were centrifuged at 18,000 g and the phosphomannose isomerase (PMI) activity of the supernatants determined. PMI activity of Jensen sarcoma and Walker carcinosarcoma 256 was equivalent to that observed in kidney or brain, but was higher than the activity of muscle. Activities in primary hepatomas, induced with 3′-methyl-4-dimethylaminoazobenzene (3′-Me-DAB) and the Novikoff hepatoma were equivalent to that of normal liver. A transplanted 3′-Me-DAB hepatoma had PMI activity 2-fold higher than normal liver, while activity in liver tissue adjacent to primary hepatomas was approximately half that observed in liver. The enzyme from the Walker tumor was inhibited by o-phenanthroline, ethylenediaminotetraacetate and p-chloromercuribenzoate (CMB). CMB inhibition was partially reversed by cysteine.

In vitro Effects of Guinea Pig Serum on the Jensen, JA-1 and JA-2 Sarcomas

Summary The in vitro growth response of the Jensen sarcoma and its nutritional variants (JA-1 and JA-2) to media containing normal guinea pig serum and partially purified asparaginase and media devoid of asparagine was studied. A comparison of the morphology of the cells grown in the presence of guinea pig serum and media devoid of asparagine showed a remarkable similarity. Further, the growth characteristics of cultures exposed to guinea pig serum and partially purified asparaginase were essentially the same. These results suggested the active component of guinea pig serum was asparaginase which catalyzed the destruction of extracellular asparagine.

Effect of tumor cell fractions upon plasma iron, liver catalase and organ weights of rats.

Summary The factor (s) in Walker carcinosarcoma 256 tissue which caused a depression in plasma iron, liver catalase activity and alterations in organ weights when injected into normal rats was localized primarily in the mitochondrial fraction.

Adenylic acid and adenosine deaminase activities in rat livers during azo-dye carcinogenesis.

Summary Adenosine and adenylic acid deaminase activities were determined in precancerous livers and in primary hepatomas induced by feeding 3′-methyl-4-dimethylamino-azobenzene. The deaminase activities in precancerous livers were increased about 100% after 30 days of dye feeding compared with control livers. Activities in primary hepatomas and in the liver adjacent to primary hepatomas were essentially the same as that of precancerous livers, indicating that no additional changes occurred upon formation of neoplastic tissues.