Cell-Density Control of Amino Acid Transport in Simian Virus 40-Transformed 3T3 Cells

Piedimonte, Giuseppe; Tramacere, Mariarosaria; Borghetti, Angelo F.

doi:10.1042/bst0071137

Cited by 10 publications

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“…Figure 8a shows that all media containing glutamine over 0.25 mM allowed a comparable cell proliferation leading to a population doubling within 24 h of incubation with growth-rate quotient (Q, Piedimonte et al, 1982) ranging between 1.1 and 1.35 at 24 h, followed by: (1) growth arrest for cells matched with initial glutamine concentrations of 0.25 and 0.5 mM and (2) slowed growth for cells exposed to higher initial glutamine concentrations (with Q of 0.14, 0.15, and 0.37 for cells exposed to initial glutamine concentrations of 0.75, 1, 2 mM, respectively). In this experiment, cell proliferation stopped after 48-72 h when glutamine concentration in the medium dropped below 0.3 mM from initial values of 0.75-2 mM ( Table 3).…”

Section: Exogenous Glutamine Requirement For Survivalmentioning

confidence: 99%

Cell susceptibility to apoptosis by glutamine deprivation and rescue: Survival and apoptotic death in cultured lymphoma-leukemia cell lines

et al. 1996

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Human leukemia/lymphoma cells maintained in culture medium without provision of fresh nutrients lose viability and die by a process resembling apoptosis within a few days. Upon incubation in an FCS-supplemented RPMI 1640 medium containing 2 mM L-glutamine CEM, Namalwa, HL-60 and U937 cells, seeded at initial densities of 0.2 to 1 x 10(6) cells/ml, ceased growing within 3-5 days and progressively entered an apoptotic pathway, as assessed by nucleosomal DNA fragmentation and morphology. Both the major energy-source nutrients in the medium, glucose and glutamine, became rapidly exhausted during the incubation. Further studies were performed using CEM cells. Incubation in glutamine-free or glucose-free medium renewed every 24 h showed that glutamine deprivation is associated with cell death by apoptosis independent of energetic failure, whereas glucose deprivation is followed by rapid loss of mitochondrial function with sharp drop of intracellular ATP and cell death by necrosis. A 12-24 h incubation in glutamine-depleted medium was required to direct the cells toward the apoptotic pathway. Growth arrest followed by apoptotic death was detected in CEM cells when medium glutamine concentration remained below 0.3-0.4 mM for at least 24 h, but a reinstatement of medium glutamine to 2 mM within this period rescued the cells from growth arrest and death.

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Section: Exogenous Glutamine Requirement For Survivalmentioning

confidence: 99%

Cell susceptibility to apoptosis by glutamine deprivation and rescue: Survival and apoptotic death in cultured lymphoma-leukemia cell lines

et al. 1996

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“…The cells were maintained in Dulbecco's modified Eagle medium (D-MEM) containing penicillin (100 units ml-') and streptomycin (100 fig ml-' Cell counting and determination of cell survival Cells were detached from the substratum by trypsinisation and appropriate dilutions of the suspension were counted in a Burker haemocytometer, as described in detail elsewhere (Piedimonte et al, 1982). Cell survival was determined by cell viability and plating efficiency.…”

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confidence: 99%

Different HSP70 expression and cell survival during adaptive responses of 3T3 and transformed 3T3 cells to osmotic stress

Petronini

Alfieri²,

Angelis³

et al. 1993

Br J Cancer

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Summary Responses both to hyperosmotic stress and to heat shock were compared in 3T3 cells, spontaneously transformed cells (ST3T3) and simian virus 40-transformed cells (SV3T3). Cell adaptation to these stresses was measured in terms of surviving cell viability and plating efficiency, while their induced synthesis of stress proteins was monitored in terms of the presence of mRNA for HSP70, the pattern of polypeptides synthesised and the accumulation of HSP70 detectable by monoclonal antibodies. All three types of cells responded similarly to heat shock in their expression of HSP70 and showed no clear differences in ability to recover. In contrast, both ST3T3 and SV3T3 cells adapted more poorly and much more slowly to hyperosmotic stress (0.5 osM incubation) than did normal 3T3 cells. This different pattern of adaptation to hyperosmotic stress was parallelled by the cells' different expression of a stress protein that could not be distinguished from the heat-induced HSP70 by any of the methods listed above. In view of these findings it seems possible that hyperosmotic treatment might be useful in selectively affecting the survival of tumour cells.Previous studies of cell metabolism and proliferation showed that transformed 3T3 cells adapted much more slowly than did 3T3 cells to hyperosmotic stress (Silvotti et al., 1991 al., 1980). These cells were routinely kept in culture for up to 2 months and discarded; then fresh cultures were started again from frozen stocks. Spontaneously transformed 3T3 (ST3T3) cells were isolated in our laboratory on the basis of their loss of density control, serum dependence, ability to grow in soft agar medium, morphological modification and increased production of lactic acid.The cells were maintained in Dulbecco's modified Eagle medium (D-MEM) containing penicillin (100 units ml-') and streptomycin (100 fig ml-'

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“…Cell numbers appeared to remain similar between days 7 and 10 for both ALI groups or slightly decreased for the static submerged control group. This can potentially be attributed to: (i) the cell population reaching saturation within the scaffold, (ii) the formation of a necrotic core due to outer cell layers impeding mass transport through the scaffold, or (iii) general plateauing of proliferation after reaching a stationary density [43,44]. The effect of initial seeding density on construct remodeling warrants further investigation.…”

Section: ) Cell Growth and Proliferationmentioning

confidence: 99%

Biomimetic system design for engineering biofidelic 3-D respiratory tissuesin vitro

Poon

Zhang

Boughton

et al. 2020

Preprint

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AbstractObjectiveThe structural and functional complexity of the respiratory system present significant challenges to capturing conditions vital for maintaining phenotypic cellular functions in vitro. Here we report a unique tissue engineering system that enables respiratory constructs to be cultured under physiological loading at an air-liquid interface (ALI).MethodsThe system consists of a porous poly-e-caprolactone scaffold mounted in a well insert, which articulates via magnetic coupling with a linear actuator device to strain attached scaffolds through a sterile barrier. For proof of concept, NCI-H460 human carcinoma cells were seeded on scaffold inserts which were subjected to 5-15% cyclic tensile strain at 0.2Hz within a six well plate. The dynamic constructs were cultured at an ALI in a standard incubator for up to 10 days along with unstimulated (static) ALI and static submerged control groups.ResultsHigh (near-100%) cell seeding efficiency was achieved within the scaffold-strain device. Both dynamic and static ALI groups yielded higher cell densities compared to the submerged control for all time points. Distinctly different patterns in cellular growth and behaviour between dynamic air-liquid interface and conventional static submerged culture groups were revealed by nuclei staining, where the actuated group displayed more uniform cellular distribution throughout the construct compared to both static controls.ConclusionAir-liquid interface culture and physiological strain are important for engineering respiratory tissue models.SignificanceThe system described allows scalable and replicable culture of 3-D tissue engineered respiratory models under biologically-relevant conditions.

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Cell-Density Control of Amino Acid Transport in Simian Virus 40-Transformed 3T3 Cells

Cited by 10 publications

References 0 publications

Cell susceptibility to apoptosis by glutamine deprivation and rescue: Survival and apoptotic death in cultured lymphoma-leukemia cell lines

Cell susceptibility to apoptosis by glutamine deprivation and rescue: Survival and apoptotic death in cultured lymphoma-leukemia cell lines

Different HSP70 expression and cell survival during adaptive responses of 3T3 and transformed 3T3 cells to osmotic stress

Biomimetic system design for engineering biofidelic 3-D respiratory tissuesin vitro

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