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

Petiot, Emma; Čuperlović-Culf, Miroslava; Shen, Chun Fang; Kamen, Amine

doi:10.1016/j.vaccine.2015.05.097

Cited by 41 publications

(50 citation statements)

References 87 publications

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“…metabolism and the deprivation of serum or replacing it with synthetic molecules are constantly increasing area of interest (24). In addition, there is some cell lines that cannot be grown is suspension mode so adherent systems are only possibilities.…”

Section: Discussionmentioning

confidence: 99%

Process Development of Adenoviral Vector Production in Fixed Bed Bioreactor: From Bench to Commercial Scale

Lesch

Heikkilä

Lipponen³

et al. 2015

Human Gene Therapy

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Large-scale vector manufacturing for phase III and beyond has proven to be challenging. Upscaling the process with suspension cells is increasingly feasible, but many viral production applications are still applicable only in adherent settings. Scaling up the adherent system has proven to be troublesome. The iCELLis(®) disposable fixed-bed bioreactors offer a possible option for viral vector manufacturing in large quantities in an adherent environment. In this study, we have optimized adenovirus serotype 5 manufacturing using iCELLis Nano with a cultivation area up to 4 m(2). HEK293 cell cultivation, infection, and harvest of the virus (by lysing the cells inside the bioreactor) proved possible, reaching total yield of up to 1.6×10(14) viral particles (vp)/batch. The iCELLis 500 is designed to satisfy demand for large-scale requirements. Inoculating a large quantity of cell mass into the iCELLis 500 was achieved by first expanding the cell mass in suspension. Upscaling the process into an iCELLis 500/100 m(2) cultivation area cassette was practical and produced up to 6.1×10(15) vp. Flask productivity per cm(2) in iCELLis Nano and iCELLis 500 was in the same range. As a conclusion, we showed for the first time that iCELLis 500 equipment has provided an effective way to manufacture large batches of adenoviral vectors.

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Section: Discussionmentioning

confidence: 99%

Process Development of Adenoviral Vector Production in Fixed Bed Bioreactor: From Bench to Commercial Scale

Lesch

Heikkilä

Lipponen³

et al. 2015

Human Gene Therapy

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show abstract

“…In combination with the previous data of the gene and protein expression levels, cell growth, substance metabolism and energy metabolism, appropriate aralar1 geneexpressing cell clone shows the highest m max , maximum cell density, DLac/DGlc, DNH 3 /DGln, ATP production rate, and the lowest ROS level, improving the metabolic performance and adenovirus reproduction capacity in HEK-293 cells. 32 Therefore, the higher the aralar1 gene expression level, the greater the extra burden on HEK-293 cells. This seriously limits the increase in the ability of the HAdV reproduction.…”

Section: Effects Of Co-overexpression Of the Aralar1 Pkm2 And Pyc2 Gmentioning

confidence: 99%

Regulation of the pyruvate metabolism node by monogene and polygene engineering of HEK-293 cells

Xie

Miao

et al. 2019

RSC Adv.

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“…To study the influence of the delivery systems proposed in a cell culture and analyze whether there was any cytotoxic effect, the HEK293T cell line, a human embryonic cell line [20], was chosen. This cell line was selected because it is an established but not a tumor one.…”

Section: Cell Culture Studiesmentioning

confidence: 99%

DSPC Liposomes Improve Transport of L-cysteine and Reduce Metabolic Activity

Perrotta

Prietto

Alonso

et al. 2016

BBJ

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Aims:In this work, we developed and characterized liposomal formulations that encapsulate Lcysteine to study their further application in drug delivery and amino acid supplementation. The lipids used were 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC). Methodology: Encapsulation efficiency and amino acid release were determined. For biophysical characterization of the three formulations, the size, surface charge and surface packing were also studied. Cell viability was analyzed with MTT reagent after treatments with formulations ir order to study efficiency of these systems in induce changes in metabolism. Results: Results showed that L-cysteine interacts at the polar head level and that this interaction stabilizes the surface charge and prevents aggregation. We also determined the influence on cell metabolism in all formulations. The presence of L-cysteine in the DSPC formulation induced deeper Perrotta et al.; BBJ, 12(4): 1-11, 2016; Article no.BBJ.24723 2 changes in metabolism, evidencing that this formulation provides better transport of this amino acid. Conclusion: Liposomes developed herein are well suited for the application in the delivery of L-cysteine. Particularly, they can encapsulate nearly all the L-cysteine and can retain it for 6 hours. Also, L-cysteine stabilized liposomes, preventing their aggregation. L-cysteine encapsulated in the DSPC formulation induced deeper changes in cell metabolism, causing a decrease in metabolic activity; this was probably due to a higher entry, thus a better liposome-mediated transport. Considering that the smaller the particle, the better the circulation, we believe that the stabilization of the vesicle by L-cysteine may allow these transporters to have higher circulation times. Based on the above, we conclude that the DSPC formulation is the best suited for further application in L-cysteine delivery. Original Research Article

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

Cited by 41 publications

References 87 publications

Process Development of Adenoviral Vector Production in Fixed Bed Bioreactor: From Bench to Commercial Scale

Process Development of Adenoviral Vector Production in Fixed Bed Bioreactor: From Bench to Commercial Scale

Regulation of the pyruvate metabolism node by monogene and polygene engineering of HEK-293 cells

DSPC Liposomes Improve Transport of L-cysteine and Reduce Metabolic Activity

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