Establishment of Clonal Strains of Rat Pituitary Tumor Cells That Secrete Growth Hormone^1,2

Abstract

Three clonal strains of epithelial cells were established from a transplantable rat pituitary tumor. These strains have been serially propagated for 14-25 months. They were subcultured every 2-3 weeks. Cells of the original strain have increased by a factor of more than 10 40 . The generation times of the 3 lines were similar and ranged between 30 and 40 hr. Cells of all 3 strains synthesize growth hormone and secrete it into the culture medium. Growth hormone synthesized in vitro is indistinguishable from normal rat pituitary growth hormone as measured by micro-complement fixation and radioimmunoassay. The specific activity of growth hormone production was estimated to be 20-40 fig/ing cell nitrogen/24 hr for the most vigorous strain. There has been no decrease in the rate of cell division nor decline in growth hormone secretion since the cells were established in culture. (Endocrinology 82: 342, 1968)

Sphingosylphosphorylcholine Activates an Amiloride‐Nsensitive Na+‐H+‐Exchange Mechanism in GH4C1 Cells

“…The cells were grown in monolayer culture in Ham's F 10 Nutrient Mixture with 15 % (by vol.) horse serum and 2.5 % fetal bovine serum in a water-saturated atmosphere of 5% CO, and 95% air at 37"C, as described previously [23].…”

Evidence That the Bone Resorption-Stimulating Factor Produced by Mouse Fibrosarcoma Cells Is Prostaglandin E2

“…Culture.--The procedures used were similar to those reported in detail previously from this laboratory for the establishment of rat pituitary tumor cells in culture (7). In brief, a tumor was removed aseptically, minced with fine scissors, and the cell aggregates dispersed with Viokase and plated in plastic Petri dishes (Falcon Plastics, Division B-D Laboratories, Inc., Los Angeles, Calif.) in Ham's F 10 medium (8) supplemented with 15% horse serum and 2.5% fetal calf serum.…”

“…Cultures were incubated at 37°C in a humidified atmosphere of 5% CO2 and 95% air. The tumor cells adapted readily to the conditions of primary culture without resort to the serial culture to animal enrichment technique previously described (7). Primary cultures were subcultured by treatment with Viokase, and the cells diluted 1 : 5 to 1: 40 and replated in fresh dishes.…”

“…To demonstrate that the cells in culture (both uncloned and clonal strains) were indeed tumor cells, we harvested cells from monolayers (7,9) and injected 106 cells per animal into groups of four to six male Swiss albino mice. All injected mice developed tumors within less than 10 days.…”

A tribute to Dr. Gordon Hisashi Sato (December 24, 1927–March 31, 2017)

“…Armed with this insight, the Sato lab devised a protocol for selecting differentiated tumor cells that were able to survive and proliferate in culture; this protocol consisted of multiple cycles of alternately passaging tumor cells in vivo and in vitro. Using this approach, they isolated continuous mammalian cell lines from rodent endocrine tumors provided by Jacob Furth (Buonassisi et al 1962;Stollar et al 1964;Yasumura et al 1966;Tashjian et al 1968;Cuprak and Sato 1968) and cell lines from other differentiated tissues (Mohit and Sato 1967;Benda et al 1968;Augusti-Tocco and Sato 1969;Rosenthal et al 1970). These cell lines were the first to exhibit differentiated traits of their tissues of origin .…”