Dependence of freeze-thaw damage on growth phase and cell cycle of cultured mammalian cells

Terasima, Toyozo; Yasukawa, Mieko

doi:10.1016/0011-2240(77)90186-9

Cited by 11 publications

(2 citation statements)

References 5 publications

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“…Two other reports have also noted that growing cells are more tolerant to stress than are nongrowing cells. While Terasima and Yasukawa (40) found plateau-phase mammalian cells to be more freeze-thaw tolerant than rapidly growing cells, Frim et al (8) reported the opposite. Similarly, Mitchel and Morrison (30) found that stationary-phase yeast cells were more resistant to UV radiation than were respirofermentative cells, but the opposite has also been reported (33).…”

Section: Discussionmentioning

confidence: 99%

Role of growth phase and ethanol in freeze-thaw stress resistance of Saccharomyces cerevisiae

Lewis

Learmonth

Watson

1993

Appl Environ Microbiol

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The freeze-thaw tolerance of Saccharomyces cerevisiae was examined throughout growth in aerobic batch culture. Minimum tolerance to rapid freezing (immersion in liquid nitrogen; cooling rate, approximately 200'C min-') was associated with respirofermentative (exponential) growth on glucose. However, maximum tolerance occurred not during the stationary phase but during active respiratory growth on ethanol accumulated during respirofermentative growth on glucose. The peak in tolerance occurred several hours after entry into the respiratory growth phase and did not correspond to a transient accumulation of trehalose which occurred at the point of glucose exhaustion. Substitution of ethanol with other carbon sources which permit high levels of respiration (acetate and galactose) also induced high freeze-thaw tolerance, and the peak did not occur in cells shifted directly from fermentative growth to starvation conditions or in two respiratorily incompetent mutants. These results imply a direct link with respiration, rather than exhaustion of glucose. The role of ethanol as a cryoprotectant per se was also investigated, and under conditions of rapid freezing (cooling rate, approximately 200°C min-'), ethanol demonstrated a significant cryoprotective effect. Under the same freezing conditions, glycerol had little effect at high concentrations and acted as a cryosensitizer at low concentrations. Conversely, under slow-freezing conditions (step freezing at -20, -70, and then -196°C; initial cooling rate, approximately 3C min-'), glycerol acted as a cryoprotectant while ethanol lost this ability.Ethanol may thus have two effects on the cryotolerance of baker's yeast, as a respirable carbon source and as a cryoprotectant under rapid-freezing conditions.

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Section: Discussionmentioning

confidence: 99%

Role of growth phase and ethanol in freeze-thaw stress resistance of Saccharomyces cerevisiae

Lewis

Learmonth

Watson

1993

Appl Environ Microbiol

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“…This is achieved by optimizing freeze medium, cell density at freezing and the freezing method, but also the treatment of cells subsequent to freezing (Day and McLellan, 1995;Doyle et al, 1999;Grout et al, 1990;Mazur et al, 1972). The success of the cryopreservation is also dependent on the general fitness of the cells prior to freezing, which is reflected in their viability, and their state in the cell cycle (Fieder et al, 2005;Koch et al, 1970;Terasima and Yasukawa, 1977). The most important single factor determining the success of cryopreservation is whether or not a cell undergoes IIF during freezing.…”

Section: Introductionmentioning

confidence: 99%

Optimal conditions for freezing CHO‐S and HEK293‐EBNA cell lines: Influence of Me₂SO, freeze density, and PEI‐mediated transfection on revitalization and growth of cells, and expression of recombinant protein

Kleman

Oellers

Lüllau

2008

Biotech & Bioengineering

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To avoid the time consuming, labor intensive seed-train expansion and to improve production reliability and consistency, portions of bulk cryopreserved cells from the same cultivation can be utilized as inocula or alternatively may be used to undertake transient transfections for large-scale bioreactor production. In this study, the conditions for large-scale freezing in cryobags were optimized utilizing a design of experiment approach. We showed that relatively high density of 30-40 x 10(6) cells/mL and relatively low Me(2)SO concentrations of 5-6% in the freezing media are optimal to freeze HEK293-EBNA and CHO-S cells in a controlled manner in order to achieve high viable cell recovery and growth post-thawing. The immediate transfer of freshly thawed cells into culture medium resulted in better cell growth compared to cells that were centrifuged in order to remove Me(2)SO. This was the case as long as the residual Me(2)SO did not exceed 0.2-0.3%. The best time to perform transient 25 kDa polyethylenimine-mediated transfection of pCEP4-EGFP plasmid into freshly thawed, one-step inoculated cells is after 72-96 h in culture. At this time point, the numbers of EGFP-positive cells in the freshly thawed culture mimic perfectly that of cells grown continuously. Finally, our data showed that it is possible to freeze transiently polyethyleneimine-transfected HEK293-EBNA cells and maintain growth rate and expression of recombinant protein following thawing. The optimal time point for freezing cells was 4 h after transfection.

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Cryopreservation and Banking of Cell Lines

Stacey

Hawkins

Fleck

2011

Animal Cell Culture

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Dependence of freeze-thaw damage on growth phase and cell cycle of cultured mammalian cells

Cited by 11 publications

References 5 publications

Role of growth phase and ethanol in freeze-thaw stress resistance of Saccharomyces cerevisiae

Role of growth phase and ethanol in freeze-thaw stress resistance of Saccharomyces cerevisiae

Optimal conditions for freezing CHO‐S and HEK293‐EBNA cell lines: Influence of Me₂SO, freeze density, and PEI‐mediated transfection on revitalization and growth of cells, and expression of recombinant protein

Cryopreservation and Banking of Cell Lines

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Dependence of freeze-thaw damage on growth phase and cell cycle of cultured mammalian cells

Cited by 11 publications

References 5 publications

Role of growth phase and ethanol in freeze-thaw stress resistance of Saccharomyces cerevisiae

Role of growth phase and ethanol in freeze-thaw stress resistance of Saccharomyces cerevisiae

Optimal conditions for freezing CHO‐S and HEK293‐EBNA cell lines: Influence of Me2SO, freeze density, and PEI‐mediated transfection on revitalization and growth of cells, and expression of recombinant protein

Cryopreservation and Banking of Cell Lines

Contact Info

Product

Resources

About

Optimal conditions for freezing CHO‐S and HEK293‐EBNA cell lines: Influence of Me₂SO, freeze density, and PEI‐mediated transfection on revitalization and growth of cells, and expression of recombinant protein