Optimization of HEK-293S cell cultures for the production of adenoviral vectors in bioreactors using on-line OUR measurements

Gálvez, J.; Lecina, Martí; Solá, Carles Miralles; Cairó, Jordi J.; Gòdia, Francesc

doi:10.1016/j.jbiotec.2011.11.007

Cited by 36 publications

(25 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As indicated in Table 2, CDE is generally observed for HEK293 cells between 1 and 2 × 10 6 cells/ml [1,61,70], whereas cell densities of 8-10 × 10 6 cells/ml can be achieved in batch cultures [64]. CDE is virus dependent with specific adenovirus production decreasing at densities higher than 2 × 10 6 cell/ml [1,70,71], while for influenza virus, high productivity is maintained up to 4 × 10 6 cells/ml in batch cultures [58,64]. Consequently, most of the transfection/infection steps are operated at cell concentrations below 4 × 10 6 cell/ml.…”

Section: Feeding Strategies and Medium Modification For Viral Productmentioning

confidence: 94%

Influence of HEK293 metabolism on the production of viral vectors and vaccine

Petiot

Čuperlović-Culf

Shen

et al. 2015

Vaccine

View full text Add to dashboard Cite

Section: Feeding Strategies and Medium Modification For Viral Productmentioning

confidence: 94%

Influence of HEK293 metabolism on the production of viral vectors and vaccine

Petiot

Čuperlović-Culf

Shen

et al. 2015

Vaccine

View full text Add to dashboard Cite

“…The medium is removed through the OT, leaving the cell spheroids in culture, until the medium level falls below the bottom of the OT. This method of volume control with continuous feeding is used in many large-scale culture applications used to generate cell products [34], [35]. The system is designed to be simple without the need for specialized sensors and automation systems to control pump speeds.…”

Section: Methodsmentioning

confidence: 99%

Nutrient Regulation by Continuous Feeding Removes Limitations on Cell Yield in the Large-Scale Expansion of Mammalian Cell Spheroids

et al. 2013

View full text Add to dashboard Cite

Cellular therapies are emerging as a standard approach for the treatment of several diseases. However, realizing the promise of cellular therapies across the full range of treatable disorders will require large-scale, controlled, reproducible culture methods. Bioreactor systems offer the scale-up and monitoring needed, but standard stirred bioreactor cultures do not allow for the real-time regulation of key nutrients in the medium. In this study, β-TC6 insulinoma cells were aggregated and cultured for 3 weeks as a model of manufacturing a mammalian cell product. Cell expansion rates and medium nutrient levels were compared in static, stirred suspension bioreactors (SSB), and continuously fed (CF) SSB. While SSB cultures facilitated increased culture volumes, no increase in cell yields were observed, partly due to limitations in key nutrients, which were consumed by the cultures between feedings, such as glucose. Even when glucose levels were increased to prevent depletion between feedings, dramatic fluctuations in glucose levels were observed. Continuous feeding eliminated fluctuations and improved cell expansion when compared with both static and SSB culture methods. Further improvements in growth rates were observed after adjusting the feed rate based on calculated nutrient depletion, which maintained physiological glucose levels for the duration of the expansion. Adjusting the feed rate in a continuous medium replacement system can maintain the consistent nutrient levels required for the large-scale application of many cell products. Continuously fed bioreactor systems combined with nutrient regulation can be used to improve the yield and reproducibility of mammalian cells for biological products and cellular therapies and will facilitate the translation of cell culture from the research lab to clinical applications.

show abstract

“…Several studies (Smith et al 1991;Hiller et al 1993;Zhang et al 1993;Kyung et al 1994) were published in the early 1990s using cellulose ester HF achieving varying densities of hybridoma or HEK293 cells from a few millions cells/mL to a maximum of 10 8 cells/mL by Kyung. Several years later, Cortin et al (2004) and Galvez et al (2012) successfully produced adenovirus in HEK 293 cell system. In 2011, Clincke et al published a comparative study of perfusion in wave bioreactor Fig.…”

Section: Tffmentioning

confidence: 99%

“…This approach requires that the cell specific glucose consumption rate is constant so it should be applied when steady-state culture is established. The oxygen uptake rate (OUR) has also been used to control the perfusion rate, given that the OUR is a good indicator of the physiological state of the cells (Kyung et al 1994;Galvez et al 2012). …”

Section: Perfusion Rate Strategy Based On Main Substrate Measurementmentioning

confidence: 99%

Perfusion Processes

Chotteau

2014

Cell Engineering

View full text Add to dashboard Cite

The interest for perfusion is increasing nowadays. This new focus has emerged from a synergy of a demand for disposable equipment and the availability of robust cell separation device, as well as the need for higher flexibility and lower investment cost. The cell separation devices mostly used today are based on filtration, i.e. alternating flow filtration, tangential flow filtration, spin-filter, or acceleration/gravity, i.e. inclined settler, centrifuge, acoustic settler. This paper gives an introduction to the basic concepts of perfusion and its practical implementation. It reviews the actual cell separation devices and describes the approaches used in the field to develop and optimize the perfusion processes.

show abstract

Optimization of HEK-293S cell cultures for the production of adenoviral vectors in bioreactors using on-line OUR measurements

Cited by 36 publications

References 26 publications

Influence of HEK293 metabolism on the production of viral vectors and vaccine

Influence of HEK293 metabolism on the production of viral vectors and vaccine

Nutrient Regulation by Continuous Feeding Removes Limitations on Cell Yield in the Large-Scale Expansion of Mammalian Cell Spheroids

Perfusion Processes

Contact Info

Product

Resources

About