Density dependent regulation of growth in suspension cultures of L‐929 cells

Glinos, André D.; Werrlein, Robert J.

doi:10.1002/jcp.1040790109

Cited by 32 publications

(11 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The growth-related regulation of AChE in WRL-10A fibroblasts raised the question of whether similar regulation of AChE occurs in other cells of fibroblastic origin, such as the 3T3 mouse and WI-38 human embryonic fibroblasts, when the growth is arrested in attached cultures upon reaching confluence. We found that under these conditions 3T3 Swiss mouse and WI-38 human fibroblasts exhibited only trace or minimal AChE activities, while WRL-10A cells, the growth of which is only minimally inhibited in attached cultures (11), showed a small AChE increase (approximately l U/rag protein), as one might expect. American Type Culture Collection.…”

Section: Distribution Of Ache a Ctivity In Other Fibroblastic Linessupporting

confidence: 66%

“…Samples from low density exponentially growing populations were obtained from cultures after exponential growth for at least 3 days. Samples from the high density populations were obtained at different times over a period of 10 40 days after establishment of the high density populations (11). Data represent the average of three determinations.…”

Section: Distribution Of Ache a Ctivity In Other Fibroblastic Linesmentioning

confidence: 99%

See 1 more Smart Citation

Properties of growth-related acetylcholinesterase in a cell line of fibroblastic origin.

Bartos¹,

Glinos²

1976

The Journal of Cell Biology

View full text Add to dashboard Cite

We have previously reported the presence and regulation of an acetylcholinehydrolyzing enzyme in high density suspension cultures of WRL-10A fibroblasts where its activity increases 100-fold when growth is arrested. Substrate specificity, substrate inhibition, and product identification studies indicate that this enzyme is acetylcholinesterase (ACHE, EC 3.1.1.7). Treatment of whole cells with 5 mM diazotized sulfanilic acid revealed that most of the AChE is located on the external surface of the cell membrane. It was also found that the enzyme is released in the medium at a rate of 0.5 U/h/mg cell protein and that within a 24-h period the de novo synthesized and liberated AChE is equivalent to 90% of the activity associated with the cells. No similar synthesis of AChE was found in six other fibroblastic cell lines examined. These and related findings indicating that acetylcholine is also present in high density populations of WRL-10A cells suggest that this unique phenotype may be used profitably in exploring further the relationship between components of the cholinergic system and non-neuronal cell growth.Growth-inhibited dense attached cultures of normal euploid mammalian cells and some established cell lines are characterized by a marked depression of total protein synthesis (24, 42, 45), while the synthesis of specialized proteins such as collagen (15,23,26,34), "SI00-protein" (33), and certain enzymes (6, 16, 35, 37) is increased.We have previously reported that densitydependent regulation of growth comparable to that of attached cell populations occurs also in suspension cultures of WRL-10A cells (10, I1, 44), a subline of L-929 mouse fibroblasts (9). In the search for additional criteria for evaluating increased synthesis of specialized proteins in this system, acetylcholinesterase (ACHE) was considered because, besides there being a description of such an increase in neuroblastoma cells by Blume et al. (5), there is suggestive evidence that the activity of this enzyme is inversely related to the rate of cell division in attached cultures of cells of mesenchymal origin as well (14), and that small but quantifiable amounts of AChE are present in L-929 cells (1,28,49). In a previous paper we demonstrated that density-dependent metabolic regulation in suspension cultures of WRL-10A cells does indeed extend to the activity of ACHE, which was found to be approximately 100-fold greater in high density growth-inhibited populations than in low density exponentially growing cultures (8).

show abstract

Section: Distribution Of Ache a Ctivity In Other Fibroblastic Linessupporting

confidence: 66%

Section: Distribution Of Ache a Ctivity In Other Fibroblastic Linesmentioning

confidence: 99%

Properties of growth-related acetylcholinesterase in a cell line of fibroblastic origin.

Bartos¹,

Glinos²

1976

The Journal of Cell Biology

View full text Add to dashboard Cite

show abstract

“…Glinos and Werrlein (1972) reported that, following dilution of a DI fibroblast culture, there was a marked, progressive increase in mean cell volume, which reached its maximum after 20 hr and then declined. These results confirm previous findings on other cell types.…”

Section: Discussionmentioning

confidence: 99%

“…Differences in mean cell size, arising during nonsynchronized growth in suspension culture, have been reported (Glinos & Werrlein, 1972), L-929 ceils from FG cultures being of larger mean size than DI cells. Curtis, personal communication).…”

mentioning

confidence: 98%

Cell size and mutual cell adhesion

1976

View full text Add to dashboard Cite

HeLa cells harvested from density-inhibited or fast growing suspension cultures, were incubated in NaCl solutions of different tonicity. Cell size enlargement produced by hypotonicity is accompanied by an increased sedimentation rate of the density-inhibited cells, whereas no appreciable change is observed in the sedimentation rate of fast growing cells. Hypotonicity also has no effect on the sedimentation rate of density-inhibited cells which previously had been treated with neuraminidase or trypsin. It is shown that the effect of hypotonicity on density-inhibited cells cannot be ascribed to release of cell surface sialic acids during hypotonic incubation. Several arguments are presented which indicate that the changes in sedimentation rate, as measured in the rotating suspension system, are not the direct consequence of the alterations in cell size, but rather must be attributed to differences in intercellular adhesiveness resulting from the size alterations. Analogous changes in intercellular adhesiveness and cell size are shown to occur during growth in isotonic suspension culture. The results can be explained by assuming that changes in cell size affect the intercellular adhesiveness by modifying the extent to which cell surface sialic acids counteract adhesion.

show abstract

“…Finally, the hypothesis focuses attention on uptake mechanisms as possibly being the primary growth regulatory mechanism in mammalian cells in vivo, emphasizes the importance of studies of these uptake mechanisms, as well as of studies of the changes in uptake that are known to take place in malignant cells (15)(16)(17)(18)(19)(20), and predicts that in instances in which the level of nutrients can be manipulated it should be possible (21) to arrest growth of malignant cells in the G1 phase of the cell cycle by limiting some critical nutrient. In particular, the suggestive evidence (1-6, 21) that a wide variety of nutrients can have regulatory functions in mammalian cells deserves exhaustive study.…”

mentioning

confidence: 99%