Large-scale propagation of a replication-defective adenovirus vector in stirred-tank bioreactor PER.C6? cell culture under sparging conditions

The aim of this study is to prepare a model for the production of Newcastle disease virus (NDV) lentogenic F strain using cell culture in bioreactor for live attenuated vaccine preparation. In this study, firstly we investigated the growth of Vero cells in several culture media. The maximum cell number was yielded by culture of Vero cells in Dulbecco's Modified Eagle Medium (DMEM) which was1.93×106 cells/ml. Secondly Vero cells were grown in two-litre stirred tank bioreactor by using several commercial microcarriers. We achieved the maximum cell concentration about7.95×105 cells/ml when using Cytodex 1. Later we produced Newcastle Disease virus in stirred tank bioreactor based on the design developed using Taguchi L4 method. Results reveal that higher multiplicity of infection (MOI) and size of cell inoculums can yield higher virus titer. Finally, virus samples were purified using high-speed centrifugation based on3∗∗(3-1) Fractional Factorial Design. Statistical analysis showed that the maximum virus titer can be achieved at virus sample concentration of 58.45% (v/v), centrifugation speed of 13729 rpm, and centrifugation time of 4 hours. As a conclusion, high yield of virus titer could be achieved through optimization of cell culture in bioreactor and separation by high-speed centrifugation.

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Ginsenoside content in suspension cultures of Panax quinquefolium L. cultivated in shake flasksand stirred-tank bioreactor

Annales Universitatis Mariae Curie-Sklodowska sectio C – Biologia ◽

10.17951/c.2017.72.1.15-26 ◽

2018 ◽

Vol 72 (1) ◽

pp. 15 ◽

Cited By ~ 1

Author(s):

Ewa Kochan ◽

Sylwia Caban ◽

Grażyna Szymańska ◽

Piotr Szymczyk ◽

Anna Lipert ◽

...

Keyword(s):

Large Scale ◽

Raw Materials ◽

Scale Up ◽

Fold Increase ◽

Suspension Cultures ◽

Stirred Tank ◽

Small Scale ◽

Panax Quinquefolium ◽

Stirred Tank Bioreactor ◽

Shake Flasks

<p>Plant suspension cultures are described as a source for the acquisition of medicinal secondary metabolites which in the future may become an alternative to traditional raw materials. This study demonstrates that the cell cultures of one of the ginseng species – Panax quinquefolium L. synthesize ginsenosides, which are triterpene saponins having a multidirectional pharmacological effects. Tested suspension cultures were run on a small scale in the shake flasksand in scale up of the process in a 10-liter stirred tank. In the shake flasks,the highest biomass yield (2.28 gl-1 for dry and 33.99 gl-1 for fresh weight) was reached on day 30 of culture, and the highest content of saponins (2.66 mg g -1 dw) was determined on day 28 of culture. In the bioreactor, nearly 2.67 and 3-fold increase of respectively dry and fresh biomass was recorded in relation to the inoculum. Large-scale cultures synthesized protopanaxatriol derivatives such as Rg1 and Re ginsenosides, however, no saponins belonging to the protopanaxadiol derivatives were reported.</p>

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Kinetic modeling and sensitivity analysis of inulinase production in large-scale stirred tank bioreactor with sugar beet molasses-based medium

Biochemical Engineering Journal ◽

10.1016/j.bej.2021.108201 ◽

2021 ◽

pp. 108201

Author(s):

Mustafa Germec ◽

Irfan Turhan

Keyword(s):

Sensitivity Analysis ◽

Sugar Beet ◽

Kinetic Modeling ◽

Large Scale ◽

Stirred Tank ◽

Stirred Tank Bioreactor ◽

Beet Molasses ◽

Sugar Beet Molasses

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Optimization of Alginante-poly-L-lysine Microencapsules Strength by Box-Behnken Model for CHO cells Culture in Stirred Tank Bioreactor

10.1101/165464 ◽

2017 ◽

Author(s):

Yu Wang ◽

Na Li ◽

Dongsheng Sun ◽

Shengnan He ◽

Lisha Mou

Keyword(s):

Cell Culture ◽

Cho Cells ◽

Protein Production ◽

Maximum Strength ◽

Stirred Tank ◽

Process Conditions ◽

Membrane Thickness ◽

Reaction Conditions ◽

Stirred Tank Bioreactor

Summary statementBox-Behnken model is an efficient method to optimized process conditions of microencapsules strength for long-term cell culture, promotes CHO cells viability and protein production in a stirred tank bioreactor.ABSTRACTCell microencapsulation technology has been proved to be a valuable technology in the fields of large-scale cell culture. It is important to accurately construct the microcapsule membranes with desired properties including a certain thickness suitable for cell growth, and maximum strength for the stability of microencapsules. As single factor experiments are time-consuming to obtain the desired membrane preparation conditions, Box-Behnken model was used to investigate the interactions among reaction conditions, predict the optimized reaction conditions for given purpose, that was membrane with maximum strength and desired thickness for microencapsulated cell culture. Significant values of R2 in this study indicated the model theoretical values are very close to the measured values. Based on the desired membrane thickness, the process of maximum strength was optimized, and the prediction agreement of measured value and model theoretical value was 91.11%. The optimized microencapsules with maximum strength and 15 μm membrane thickness promote CHO cells viability and protein production in stirred tank bioreactor. The result shows that Box-Behnken model is an efficient method to optimized process conditions of microencapsules strength for long-term cell culture.

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