Cryopreservation of Animal and Human Cell Lines

Morris, Christopher B.

doi:10.1385/0-89603-296-5:179

Cited by 7 publications

(8 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Optimization experiments for freezer profile development focused on adjusting the controlled freezing parameters (heat of fusion hold temperature and time), for various FROSTI rack positions (using a custom 36‐slot rack) inside the controlled rate freezer to minimize variation in temperature profiles and minimize deviation from the target cooling rate. In order to minimize detrimental effects on post‐thaw viability of CHO cells, the rate of cooling is controlled between 1 and 5°C/min until the temperature is below −40°C, at which point the cells have frozen internally (Morris, 2007).…”

Section: Resultsmentioning

confidence: 99%

“…The impact of freezing CHO cells at varying cell densities ranging from ∼30 × 10 6 cells/mL (low density, LD), ∼60 × 10 6 cells/mL (mid density, MD), and, ∼110 × 10 6 cells/mL (high density, HD) on post‐thaw performance was evaluated in a study performed for three CHO Cell Lines B, C, and D. Post‐thaw performance was evaluated in 2‐L bioreactors, and the results are presented in Figure 5A and B. The low‐density case corresponds to the typical freezing density of ∼30 × 10 6 cells/mL in cell bank ampoules (Morris, 2007), and can be considered as a control for each of these comparisons.…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Development of a new bioprocess scheme using frozen seed train intermediates to initiate CHO cell culture manufacturing campaigns

Seth

Hamilton

Stapp

et al. 2013

Biotech & Bioengineering

View full text Add to dashboard Cite

Agility to schedule and execute cell culture manufacturing campaigns quickly in a multi-product facility will play a key role in meeting the growing demand for therapeutic proteins. In an effort to shorten campaign timelines, maximize plant flexibility and resource utilization, we investigated the initiation of cell culture manufacturing campaigns using CHO cells cryopreserved in large volume bags in place of the seed train process flows that are conventionally used in cell culture manufacturing. This approach, termed FASTEC (Frozen Accelerated Seed Train for Execution of a Campaign), involves cultivating cells to high density in a perfusion bioreactor, and cryopreserving cells in multiple disposable bags. Each run for a manufacturing campaign would then come from a thaw of one or more of these cryopreserved bags. This article reviews the development and optimization of individual steps of the FASTEC bioprocess scheme: scaling up cells to greater than 70 × 10(6) cells/mL and freezing in bags with an optimized controlled rate freezing protocol and using a customized rack configuration. Flow cytometry analysis was also employed to understand the recovery of CHO cells following cryopreservation. Extensive development data were gathered to ensure that the quantity and quality of the drug manufactured using the FASTEC bioprocess scheme was acceptable compared to the conventional seed train process flow. The result of offering comparable manufacturing options offers flexibility to the cell culture manufacturing network.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Development of a new bioprocess scheme using frozen seed train intermediates to initiate CHO cell culture manufacturing campaigns

Seth

Hamilton

Stapp

et al. 2013

Biotech & Bioengineering

View full text Add to dashboard Cite

show abstract

“…Another interesting point that emerged from our study was that the thawing rate impacted significantly on post-thaw semen quality and fertilizing capacity. In this regard, it is known that the thawing rate is among the most critical factors that influence sperm frozen cryosurvival [11,52,53], other than the fact that it is also the most sensitive parameter in the cryopreservation of Salmonidae semen [54,55]. In particular, the lower thawing rate (10 °C for 30 s) recorded better post-thaw semen quality for all NP-CPAs, regardless of the concentrations used.…”

Section: Discussionmentioning

confidence: 99%

Optimization of Sperm Cryopreservation Protocol for Mediterranean Brown Trout: A Comparative Study of Non-Permeating Cryoprotectants and Thawing Rates In Vitro and In Vivo

Rusco

Iorio

Gibertoni³

et al. 2019

Animals

View full text Add to dashboard Cite

The aim of our study was to test the effects of different non-permeating cryoprotectants (NP-CPAs), namely low-density lipoproteins (LDLs), sucrose, and egg yolk, and thawing rates on the post-thaw semen quality and fertilizing ability of the native Mediterranean brown trout. Pooled semen samples were diluted 1:3 (v:v) with 2.5%, 5%, 10%, or 15% LDL; 0.05, 0.1, or 0.3 M sucrose; or 10% egg yolk. At the moment of analysis, semen was thawed at 30 °C/10 s or 10 °C/30 s. The post-thaw semen quality was evaluated, considering motility, the duration of motility, viability, and DNA integrity. Significantly higher values of motility and viability were obtained using egg yolk/10 °C for 30 s, across all treatments. However, LDL and sucrose concentrations affected sperm cryosurvival, showing the highest post-thaw sperm quality at 5% LDL and 0.1 M sucrose. Based on the in vitro data, egg yolk, 5% LDL, and 0.1 M sucrose thawed at 10 °C or 30 °C were tested for the in vivo trial. The highest fertilization and hatching rates were recorded using egg yolk/10 °C (p < 0.05). According to these in vitro and in vivo results, egg yolk emerged as the most suitable NP-CPA and 10 °C/30 s as the best thawing rate for the cryopreservation of this trout sperm, under our experimental conditions.

show abstract

“…Aliquots (1 mL) were then transferred to pre-labelled cryovials. A two-step cryopreservation protocol based on that established by Morris ( 1981 ), cooling at 1 °C minute −1 to −40 °C, with the addition of automated ice-nucleation at −5 °C was employed. After being held for 15 min at −40 °C, all samples were plunged into liquid nitrogen.…”

Section: Methodsmentioning

confidence: 99%

Cryopreservation studies of an artificial co-culture between the cobalamin-requiring green alga Lobomonas rostrata and the bacterium Mesorhizobium loti

2017

View full text Add to dashboard Cite

Algal-bacterial co-cultures, rather than cultures of algae alone, are regarded as having the potential to enhance productivity and stability in industrial algal cultivation. As with other inocula in biotechnology, to avoid loss of production strains, it is important to develop preservation methods for the long-term storage of these cultures, and one of the most commonly used approaches is cryopreservation. However, whilst there are many reports of cryopreserved xenic algal cultures, little work has been reported on the intentional preservation of both algae and beneficial bacteria in xenic cultures. Instead, studies have focused on the development of methods to conserve the algal strain(s) present, or to avoid overgrowth of bacteria in xenic isolates during the post-thaw recovery phase. Here, we have established a co-cryopreservation method for the long-term storage of both partners in a unialgal-bacterial co-culture. This is an artificial model mutualism between the alga Lobomonas rostrata and the bacterium Mesorhizobium loti, which provides vitamin B12 (cobalamin) to the alga in return for photosynthate. Using a Planer Kryo 360 controlled-rate cooler, post-thaw viability (PTV) values of 72% were obtained for the co-culture, compared to 91% for the axenic alga. The cultures were successfully revived after 6 months storage in liquid nitrogen, and continued to exhibit mutualism. Furthermore, the alga could be cryopreserved with non-symbiotic bacteria, without bacterial overgrowth occurring. It was also possible to use less controllable passive freezer chambers to cryopreserve the co-cultures, although the PTV was lower. Finally, we demonstrated that an optimised cryopreservation method may be used to prevent the overgrowth potential of non-symbiotic, adventitious bacteria in both axenic and co-cultures of L. rostrata after thawing.

show abstract

Cryopreservation of Animal and Human Cell Lines

Cited by 7 publications

References 0 publications

Development of a new bioprocess scheme using frozen seed train intermediates to initiate CHO cell culture manufacturing campaigns

Development of a new bioprocess scheme using frozen seed train intermediates to initiate CHO cell culture manufacturing campaigns

Optimization of Sperm Cryopreservation Protocol for Mediterranean Brown Trout: A Comparative Study of Non-Permeating Cryoprotectants and Thawing Rates In Vitro and In Vivo

Cryopreservation studies of an artificial co-culture between the cobalamin-requiring green alga Lobomonas rostrata and the bacterium Mesorhizobium loti

Contact Info

Product

Resources

About