scholarly journals Upstream process optimization and micro- and macrocarrier screening for large-scale production of the oncolytic H-1 protoparvovirus

2021 ◽  
Author(s):  
Daniel Wohlfarth ◽  
Veronika Frehtman ◽  
Marcus Müller ◽  
Martin Vogel ◽  
Linh Minh Phuc Phan ◽  
...  
Keyword(s):  
Cell Culture ◽  
Production Process ◽  
Large Scale ◽  
Culture Medium ◽  
Feeding Strategy ◽  
Virus Yield ◽  
Seeding Density ◽  
Scale Production ◽  
Cell Culture Medium ◽  
Transfer Capability

Abstract The oncolytic virus H-1PV is a promising candidate for various cancer treatments. Therefore, production process needs to be optimized and scaled up for future market release. Currently, the virus is produced with minimum essential medium in 10-layer CellSTACK® chambers with limited scalability, requiring a minimum seeding density of 7.9E3 cells/cm2. Production also requires a 5% fetal bovine serum (FBS) supplementation and has a virus yield up to 3.1E7 plaque-forming units (PFU)/cm2. Using the animal-free cell culture medium VP-SFM™ and a new feeding strategy, we demonstrate a yield boost by a mean of 0.3 log while reducing seeding density to 5.0E3 cells/cm2 and cutting FBS supplementation by up to 40% during the production process. Additionally, FBS is completely removed at the time of harvest. Eleven commercial micro- and macrocarriers were screened regarding cell growth, bead-to-bead transfer capability, and virus yield. We present a proof-of-concept study for producing H-1PV on a large scale with the microcarrier Cytodex® 1 in suspension and a macrocarrier for a fixed-bed iCELLis® bioreactor. A carrier-based H-1PV production process combined with an optimized cell culture medium and feeding strategy can facilitate future upscaling to industrial-scale production. Key points • Virus yield increase and FBS-free harvest after switching to cell culture medium VP-SFM™. • We screened carriers for cell growth, bead-to-bead transfer capability, and H-1PV yield. • High virus yield is achieved with Cytodex® 1 and macrocarrier for iCellis® in Erlenmeyer flasks.

scholarly journals A Novel Approach Using Conventional Methodologies to Scale up BNC Production Using Komagataeibacter medellinensis and Rotten Banana Waste as Alternative

Processes ◽  
2020 ◽  
Vol 8 (11) ◽  
pp. 1469
Author(s):  
Carlos Molina-Ramírez ◽  
Juan Álvarez ◽  
Robin Zuluaga ◽  
Cristina Castro ◽  
Piedad Gañán
Keyword(s):  
Production Process ◽  
Large Scale ◽  
Culture Medium ◽  
Scale Up ◽  
Glucose Consumption ◽  
Human Consumption ◽  
Scale Production ◽  
Bacterial Nanocellulose ◽  
Large Scale Production ◽  
Glucose Diffusion

Currently, cellulose nanostructures are among the most promising structures, and extensive work in materials and biotechnology industries is aimed at identifying an efficient process of production. Even when production at the laboratory scale is successful, crucial aspects of increased commercial applications for cellulose nanostructures are linked to large-scale production. Large-scale production requires a balance between the cost of the culture medium and product value. Therefore, in this work, for the optimization and scaling up of bacterial nanocellulose, a culture medium consisting of rotten banana unsuitable for human consumption was used for the first time as an inexpensive feedstock. Initially, the bacterial nanocellulose (BNC) culture medium conditions were optimized, and it was established that a glucose concentration of 26.4 g/L and a V/A ratio of 2.2 cm were the optimal conditions for production reaching a BNC yield of 5 g/L, which was 42.4% higher than the best result initially obtained. Finally, the scale-up process was performed, implementing a regime analysis methodology by comparing the characteristic times of the critical mechanisms involved in BNC production, namely, microbial growth, glucose consumption, BNC production, and glucose diffusion into the BNC membrane, as the first approach for this type of BNC production process. The mechanism underlying the BNC production process is glucose diffusion into the BNC membrane (characteristic time, 675.47 h). Thus, the V/A ratio was selected as the scale-up criterion most suitable for producing BNC under static culture conditions, allowing the production of 16 g of BNC after 12 d of fermentation in a plastic bioreactor, which was 3378% higher than that produced in glass vessels. The results obtained in this study may initiate further improvements in BNC commercial production by exploiting different feedstocks.

3-iodothyronamine (T1AM) is taken up and rapidly metabolized in cell culture medium only at low concentration

2020 ◽  
Author(s):  
Federica Saponaro ◽  
Marco Borsò ◽  
Sara Verlotta ◽  
Lavinia Bandini ◽  
Alessandro Saba ◽  
...  
Keyword(s):  
Cell Culture ◽  
Culture Medium ◽  
Cell Culture Medium ◽  
Low Concentration

Effect of Atmospheric-Pressure Helium Plasma Jet on Cell Culture Medium

2013 ◽  
Vol 133 (5) ◽  
pp. 278-285
Author(s):  
Norimitsu Takamura ◽  
Douyan Wang ◽  
Takao Satoh ◽  
Takao Namihira ◽  
Hisato Saitoh ◽  
...  
Keyword(s):  
Cell Culture ◽  
Atmospheric Pressure ◽  
Culture Medium ◽  
Plasma Jet ◽  
Helium Plasma ◽  
Cell Culture Medium

scholarly journals Defined MSC exosome with high yield and purity to improve regenerative activity

2021 ◽  
Vol 12 ◽  
pp. 204173142110086
Author(s):  
Jun Yong Kim ◽  
Won-Kyu Rhim ◽  
Yong-In Yoo ◽  
Da-Seul Kim ◽  
Kyoung-Won Ko ◽  
...  
Keyword(s):  
Cell Culture ◽  
Culture Medium ◽  
Density Lipoprotein ◽  
High Yield ◽  
Human Umbilical Cord ◽  
Cell Culture Medium ◽  
Human Coronary Artery ◽  
Isolation Methods ◽  
Regenerative Activity ◽  
Yield And Purity

Exosomes derived from mesenchymal stem cells (MSCs) have been studied as vital components of regenerative medicine. Typically, various isolation methods of exosomes from cell culture medium have been developed to increase the isolation yield of exosomes. Moreover, the exosome-depletion process of serum has been considered to result in clinically active and highly purified exosomes from the cell culture medium. Our aim was to compare isolation methods, ultracentrifuge (UC)-based conventional method, and tangential flow filtration (TFF) system-based method for separation with high yield, and the bioactivity of the exosome according to the purity of MSC-derived exosome was determined by the ratio of Fetal bovine serum (FBS)-derived exosome to MSC-derived exosome depending on exosome depletion processes of FBS. The TFF-based isolation yield of exosome derived from human umbilical cord MSC (UCMSC) increased two orders (92.5 times) compared to UC-based isolation method. Moreover, by optimizing the process of depleting FBS-derived exosome, the purity of UCMSC-derived exosome, evaluated using the expression level of MSC exosome surface marker (CD73), was about 15.6 times enhanced and the concentration of low-density lipoprotein-cholesterol (LDL-c), known as impurities resulting from FBS, proved to be negligibly detected. The wound healing and angiogenic effects of highly purified UCMSC-derived exosomes were improved about 23.1% and 71.4%, respectively, with human coronary artery endothelial cells (HCAEC). It suggests that the defined MSC exosome with high yield and purity could increase regenerative activity.

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