Morphologic and biochemical variability of tissue and cultured cells from human pheochromocytoma

Abstract

Primary cell cultures from 18 human pheochromocytomas were maintained in culture for 10 to 12 days and characterized. The cell yields ranged from 1.0 to 60.1 X 10(6) cells/g wet weight of tissue. Cell size, as determined by histofluorescent microscopy, varied as much as seven-fold among cells derived from a given tumor and ten-fold between cells from all tumors. Cell catecholamine content, norepinephrine (NE) plus epinephrine, ranged from 0.4 to 89.5 nmol/10(6) cells at day 5 in culture and did not correlate with catecholamine content of the tissue from which the cells were obtained. Cell catecholamine content decreased with time in culture, but this decrease could not be related to a change in cell viability, the type of media used, an inability to convert dopamine to NE, or an alteration in the uptake of 3H-NE. Cellular uptake of 1.0 microM 3H-NE varied as much as 230-fold between all cell dispersions. The basal and acetylcholine stimulated release of both preloaded 3H-NE and the endogenous catecholamines was quite variable. There was no correlation between the release rate, either basal or stimulated, of preloaded 3H-NE and the endogenous catecholamines. This study represents the largest existing data base on culturing cells from these tumors and describes many of the morphologic and biochemical characteristics of this cell system.

Possible autocrine enkephalin regulation of catecholamine release in human pheochromocytoma cells

“…In addition, and in a similar way to that previously described by other authors (Tischler et al, 1984;Jaques and Tobes, 1986), we have observed a decrease in catecholamine levels with the progression of the culture. The decrease in norepinephrine, epinephrine and dihydroxyphenylglycol observed in cell cultures does not seem to be related to a dedifferentiation process as catecholamine precursors, DOPA and dopamine did not increase concomitantly.…”

“…As for the cell content in culture with the passing of time, results shown in the literature are somehow contradictory. On the one hand a marked loss of cells after 21 days in culture has been demonstrated (Tischler et al,1984) while on the other no loss of cells over a period of 12 days in culture has been shown (Jaques and Tobes, 1986). In our experimental conditions -5 days in culture -no change in cell content was observed.…”

“…Several authors have shown that cells obtained from human pheochromocytoma are readily recognized as polygonal, dark-phase epithelial-like cells that usually form clusters and give rise to thin, branching, varicose processes of varying degrees similar to those of cultured neurons (Tischler et al, 1976(Tischler et al, , 1984Jaques and Tobes, 1986). Even though we used a different isolation protocol, similar cell populations to the ones described by these authors were obtained: a) cells firmly attached to the surface of the plate, with branched processes appearing with the progression of the culture, and b) round dark-phase cells in suspension.…”

“…Few in vitro studies are available and the existing ones mainly describe catecholamine content of human pheochromocytoma cells after a long period in culture. In those studies, a loss or alteration of storage granules (Misugi et al, 1968;Jarry et al, 1989) and a decrease in catecholamine content (Tischler et al, 1984;Jaques and Tobes, 1986) have been observed with the passing of time. Evidence about hormonal regulation of catecholamine production by human pheochromocytoma cells in culture is scarce and it has also been obtained in long-term cultures.…”