Regulation of protein secretion in Chinese hamster ovary cells by cell cycle position and cell density *1Plasminogen activator, procollagen and fibronectin

Aggeler, Judith; Kapp, Leon N.; Tseng, Scheffer C.G.; Werb, Zena

doi:10.1016/0014-4827(82)90252-x

Cited by 47 publications

(26 citation statements)

References 18 publications

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“…It is also in contrast to the findings of Kubbies et al (1990) and Scott et al (1987) who found a direct correlation of tPA production with the S phase. Finally, Aggeler et al (1982) observed peak tPA production during the G2/M phases. Of course, in all these cases, the confounding factor remains consistency of analysis and interpretation of the experimental data.…”

Section: Dhfr Expressionmentioning

confidence: 83%

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Cell cycle phase dependent productivity of a recombinant Chinese hamster ovary cell line

Dutton¹,

Scharer

Moo-Young

2007

Cytotechnology

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A Chinese Hamster Ovary cell line, CHO1-15 500 , producing recombinant human tissue type plasminogen activator (tPA) via the dihydrofolate reductase (DHFR) amplification system, was studied in batch culture. In this system both DHFR and tPA are under the control of the strong constitutive viral SV40 early promoter. Employing the cumulative viable cell-hour approach, the specific productivity of tPA had maxima in the lag (0.065 pg cell -1 h -1 ) and early decline (0.040 pg cell -1 h -1 ) population growth phases. The viable population was assigned into four subpopulations (G1, S, G2/M phase, and Apoptotic cells) using flow cytometric analysis. As expected, intracellular DHFR was maximally expressed during the S cell cycle phase. The production of tPA, however, was found to be a direct linear function of the G1 phase, with a subpopulation specific productivity of 0.080 pg c-h -1. Productivity maxima in the lag and early decline corroborate the flow cytometric data, indicative that this recombinant tPA production occurs primarily in the G1 cell cycle phase, not the S phase. This suggests that endogenous regulatory mechanisms are important controlling influences on the production of recombinant tPA in this ovarian cell line. Productivity from recombinant cell lines cannot be inferred from either the plasmid construct or the host cell alone. Elucidation of the relationship between expression of recombinant protein and the cell cycle phases of the host cell is a major component of the characterization of the animal cell production system. This information facilitates rational process design, including operating mode, modelling and control, and medium formulation.Keywords Cell cycle associated protein productivity Á Chinese hamster ovary (CHO) Á DHFR Á Regulation of expression Á Specific productivity Á Tissue plasminogen activator (tPA) Abbreviation CH vol volumetric cell-hours

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Section: Dhfr Expressionmentioning

confidence: 83%

“…The activity of plasminogen activators has been reported to be increased in mitotic fibroblasts (Aggeler et al 1982). These workers found that plasminogen activators were secreted from fibroblasts in culture during the G2 and M phases, with the activity being about twice that associated with the early cell cycle phases.…”

Section: Dhfr Expressionmentioning

confidence: 97%

Cell cycle phase dependent productivity of a recombinant Chinese hamster ovary cell line

Dutton¹,

Scharer

Moo-Young

2007

Cytotechnology

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“…Therefore we measured the amount of fibronectin (FN) secreted into the media of CHOwt and CHO-A6 cells (Aggeler et al , 1982; Figure 10A). Consistent with AnxA6 inhibiting t-SNARE function, FN secretion was strongly reduced in CHO-A6 cells after 24 h (31%) and 48 h (53%).…”

Section: Resultsmentioning

confidence: 99%

Cholesterol transport from late endosomes to the Golgi regulates t-SNARE trafficking, assembly, and function

Reverter¹,

Rentero²,

Muga³

et al. 2011

Molecular Biology of the Cell

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This study shows that impaired cholesterol egress from late endosomes in cells with high annexin A6 levels is associated with altered soluble N-ethylmaleimide–sensitive fusion protein 23 (SNAP23) and syntaxin-4 cellular distribution and assembly and accumulation in Golgi membranes. This correlates with reduced secretion of cargo along the constitutive and SNAP23/syntaxin-4–dependent secretory pathway.

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“…The modulation of cell function induced by intercellular contacts has already been observed in cultured cells (1,12,19). Since secretion of fibronectin is stimulated by cell-to-cell contacts (2,11), secreted fibronectin may be responsible for the promotion of survival by prior growth in high-density cultures through a aynergistic interaction with fibroblast growth factor.…”

Section: Resultsmentioning

confidence: 92%

“…Some types of fibroblastic cell are known to convert to adipocytes when intercellular contacts are made (8,9). The contact-mediated modulation of cell function is also observed in various other types of cell, for example, the stimulation of secretion of fibronectin (2,11) and modulation of the organization of the cytoskeleton (27). Changes in cell function induced by intercellular contacts may modulate the ability of the cells to survive in the absence of calf serum.…”

mentioning

confidence: 99%

Survival of 3T3-L1 cells induced by fibroblast growth factor depends on cell density and adhesion to the substratum.

Shiba

Yamada

Kanno

1989

Cell Struct. Funct.

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ABSTRACT. The effects of cell contacts and the attachment of cells to the substratum on growth-factor-induced survival of 3T3-L1 cells were investigated to clarify their involvement in the maintenance of cell viability. When 3T3-L1 cells in low-density cultures or in high-density cultures were harvested with EDTA solution and cultured in the absence of calf serum, almost all cells from the low-density cultures lost viability 24 h later. However, about 15% of the cells harvested from high-density cultures survived for 24 h in the absence of calf serum. Addition of calf serum also enhanced the survival of cells from high-density cultures to a much greater extent than that of cells from low-density cultures. Addition of fibroblast growth factor enhanced the survival of cells, especially in the case of cells from high-density cultures. However, epidermal growth factor and platelet-dervied growth factor failed to enhance survival. Coating of culture dishes with vitronectin slightly enhanced cell survival. Addition of fibroblast growth factor markedly enhanced the survival of cells on the dishes coated with vitronectin or with fibronectin, but not on the dishes coated with heat-denatured bovine serum albumin. These results suggest that fibroblast growth factor promotes survival of 3T3-L1 cells, depending on cellto-cell contacts during prior culture and on the adhesion of cells to the substratum.

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Regulation of protein secretion in Chinese hamster ovary cells by cell cycle position and cell density *1Plasminogen activator, procollagen and fibronectin

Cited by 47 publications

References 18 publications

Cell cycle phase dependent productivity of a recombinant Chinese hamster ovary cell line

Cell cycle phase dependent productivity of a recombinant Chinese hamster ovary cell line

Cholesterol transport from late endosomes to the Golgi regulates t-SNARE trafficking, assembly, and function

Survival of 3T3-L1 cells induced by fibroblast growth factor depends on cell density and adhesion to the substratum.

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