The transient expression of CHIKV VLP in large stirred tank bioreactors.

Abstract:

:Transient gene expression (TGE) bioprocesses have been difficult to scale up in large stirred tank bioreactors with volumes of more than 1.5 L. Low production levels are often observed, but the causes have not been investigated (Gutierrez-Granados et al. in Crit Rev Biotechnol 38:918-940, 2018). Chikungunya Virus-like particle (VLP), expressed by DNA-PEI transient transfection, is a representative case study for these difficulties. Clinical materials were produced in shake flasks, but the process suffered when transferred to large stirred tank bioreactors. The resulting process was not operationally friendly nor cost effective. In this study, a systematic approach was used to investigate the root causes of the poor scale up performance. The transfection conditions were first screened in ambr® 15 high throughput mini bioreactors then examined in 3 L stirred-tank systems. The studies found that production level was negatively correlated with inoculum cell growth status (P < 0.05). The pH range, DNA and PEI levels, order of the reagent addition, and gas-sparging systems were also studied and found to affect process performance. Further hydromechanical characterizations (Re, energy dissipation rates, and P/V, etc.) of shake flasks, ambr® 15, and 3-L stirred tank systems were performed. Overall, the study discovered that the shear stress (caused by a microsparger) and PEI toxicity together were the root causes of scale-up failure. Once the microsparger was replaced by a macrosparger, the scale-up was successful.

journal_name

Cytotechnology

journal_title

Cytotechnology

authors

Chen P,Demirji J,Ivleva VB,Horwitz J,Schwartz R,Arnold F

doi

10.1007/s10616-019-00346-x

subject

Has Abstract

pub_date

2019-12-01 00:00:00

pages

1079-1093

issue

6

eissn

0920-9069

issn

1573-0778

pii

10.1007/s10616-019-00346-x

journal_volume

71

pub_type

杂志文章
  • Neuroscience of water molecules: a salute to professor Linus Carl Pauling.

    abstract::More than 35 years ago double Nobel laureate Linus Carl Pauling published a powerful model of the molecular mechanism of general anesthesia, generally referred to as the hydrate-microcrystal (aqueous-phase) theory. This hypothesis, based on the molecular behavior of water molecules, did not receive serious attention d...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-009-9216-x

    authors: Nakada T

    更新日期:2009-04-01 00:00:00

  • Induction of MRP/GS-X pump and cellular resistance to anticancer prostaglandins.

    abstract::We provide evidence that the expression of the human MRP/GS-X pump encoded by the MRP (multidrug resistance associated protein) gene is induced by cisplatin in human leukemia HL-60/R-CP (cisplatin-resistant) cells and modulates cell growth inhibition by delta(7)-prostaglandin A1 (PGA1) methyl ester. The MRP mRNA level...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/BF00744216

    authors: Akimaru K,Kuo MT,Furuta K,Suzuki M,Noyori R,Ishikawa T

    更新日期:1996-01-01 00:00:00

  • PLCgamma2 Activates CREB-dependent Transcription in PC12 Cells Through Phosphorylation of CREB at Serine 133.

    abstract::The cAMP and Ca(2+) signaling pathways activate the transcription factor CREB through its phosphorylation at Serine 133. Activation of CREB is involved in the regulation of various biological phenomena. To understand further the mechanisms of the regulation of CREB activity in response to activation of the cAMP and Ca...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-005-3763-6

    authors: Iwamoto T,Mamiya N,Masushige S,Kida S

    更新日期:2005-01-01 00:00:00

  • Rapid titration of adenoviral infectivity by flow cytometry in batch culture of infected HEK293 cells.

    abstract::There is a constant and growing interest in exploitingadenoviruses as vectors for gene therapy when transientexpression of a therapeutic protein is necessary. Therequirement for an increased viral titre has prompted asearch for techniques by which this virus may be assayedwith greater speed and simplicity. Conventiona...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1023/A:1021106116887

    authors: Gueret V,Negrete-Virgen JA,Lyddiatt A,Al-Rubeai M

    更新日期:2002-01-01 00:00:00

  • Synthesis, characterization and cytotoxicity of boron nitride nanoparticles: emphasis on toxicogenomics.

    abstract::Nanotechnology is increasingly developing area including more than 700 commercial products such as clothing, food preparation, cosmetics, mechanics, electronics and also health industry. People generally contact with nanoparticles by inhaling from air. Thus, it is becoming important issue to understand harmful effects...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-019-00292-8

    authors: Türkez H,Arslan ME,Sönmez E,Açikyildiz M,Tatar A,Geyikoğlu F

    更新日期:2019-02-01 00:00:00

  • Utility of NucleoCounter for the chondrocyte count in the collagenase digest of human native cartilage.

    abstract::In cartilage tissue engineering, viable cell numbers should be correctly counted in the collagenase digest of the biopsied cartilage. However, this is a difficult task due to the presence of matrix debris, cell ghosts and their aggregates. To search for the correct cell counting method in this situation, we evaluated ...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-010-9304-y

    authors: Yonenaga K,Nishizawa S,Akizawa M,Asawa Y,Fujihara Y,Takato T,Hoshi K

    更新日期:2010-12-01 00:00:00

  • Analysis of toxicity effects of Di-(2-ethylhexyl) phthalate exposure on human bronchial epithelial 16HBE cells.

    abstract::Recent studies have indicated that Di-(2-ethylhexyl) phthalate (DEHP), the most commonly used plasticizer in daily-life products, could be dispersed in indoor air and induce human exposure via inhalation. DEHP has been reported to have effects on the respiratory system in both animal and human researches. The toxicity...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-017-0111-6

    authors: Ma Y,Guo Y,Wu S,Lv Z,Zhang Q,Xie X,Ke Y

    更新日期:2018-02-01 00:00:00

  • Human fetal aorta-derived vascular progenitor cells: identification and potential application in ischemic diseases.

    abstract::Vasculogenesis, the formation of blood vessels in embryonic or fetal tissue mediated by immature vascular cells (i.e., angioblasts), is poorly understood. Here we report a summary of our recent studies on the identification of a population of vascular progenitor cells (VPCs) in human fetal aorta. These undifferentiate...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-008-9167-7

    authors: Invernici G,Madeddu P,Emanueli C,Parati EA,Alessandri G

    更新日期:2008-09-01 00:00:00

  • Alteration of mammalian cell metabolism by dynamic nutrient feeding.

    abstract::The metabolism of hybridoma cells was controlled to reduce metabolic formation in fed-batch cultures by dynamically feeding a salt-free nutrient concentrate. For this purpose, on-line oxygen uptake rate (OUR) measurement was used to estimate the metabolic demand of hybridoma cells and to determine the feeding rate of ...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1023/A:1007945826228

    authors: Zhou W,Rehm J,Europa A,Hu WS

    更新日期:1997-07-01 00:00:00

  • Immunonucleochemistry: a new method for in situ detection of antigens in the nucleus of cells in culture.

    abstract::The advancement of immunocytochemistry (ICC) allows one to observe detailed spatial distribution of cellular antigens, but, with some limitations. Using conventional ICC, it is difficult to distinguish the nuclear localization from cytoplasm, as two large subcellular compartments overlap on the z-axis. In this study, ...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-010-9266-0

    authors: Moon IS,Lee H,Park SD,Seog DH

    更新日期:2010-04-01 00:00:00

  • Localization of viral transforming sequences within marker chromosomes associated with tumor formation and progression in a murine fibrosarcoma.

    abstract::The low-metastatic RSV-transformed fibrosarcoma line B77-3T3 and its metastatic variant AA12, selected in vitro, have been analysed by blot and in situ hybridization with v-src and murine c-myc specific probes in order to detect molecular rearrangements underlying the transition from the low-metastatic to the high-met...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/BF00351122

    authors: Doneda L,Custode P,Morghen Cde G,Larizza L

    更新日期:1987-10-01 00:00:00

  • Insights into protective effects of medium additives on animal cells under fluid stresses: the hydrophobic interactions.

    abstract::Animal cells in suspension culture can suffer severe mechanical damage from bursting gas bubbles or other hydrodynamic force sources. Certain chemical additives in the culture media, particularly some surface-active chemicals, can effectively protect animal cells against such damage. Previously we proposed that the pr...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/BF00353929

    authors: Wu J

    更新日期:1996-01-01 00:00:00

  • A modification of tooth germ cultivation in vitro and in ovo.

    abstract::Mandibular molar anlages excised from 17-day mouse foetuses were cultured in vitro or in ovo (on the chorioallantoic membrane). In both cases, the explants were underlain either with a Millipore filter or with a piece of fibrin foam. Tooth germs were harvested after 7 days of cultivation and processed histologically. ...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/BF00135638

    authors: Peterka M,Mandys V,Peterková R

    更新日期:1991-09-01 00:00:00

  • Regulation of matrix metalloproteinase-1 and alpha-smooth muscle actin expression by interleukin-1 alpha and tumour necrosis factor alpha in hepatic stellate cells.

    abstract::Hepatic stellate cells (HSCs) are key players in liver fibrosis and regeneration via collagen degradation and synthesis. These phenomena involve inflammatory cytokines released from non-parenchymal liver cells such as Kupffer cells. Although the effects of individual cytokines on many cell types have been investigated...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-016-9948-3

    authors: Inoue A,Obayashi K,Sonoda Y,Nakamura A,Ueno T,Kuhara S,Tashiro K

    更新日期:2017-06-01 00:00:00

  • Silencing of soluble epoxide hydrolase 2 gene reduces H2O2-induced oxidative damage in rat intestinal epithelial IEC-6 cells via activating PI3K/Akt/GSK3β signaling pathway.

    abstract::Oxidative stress plays a vital role in the occurrence and development of intestinal injury. Soluble epoxide hydrolase 2 gene (EPHX2) is a class of hydrolytic enzymes. We aim to explore the effects and molecular mechanism of siEPHX2 on H2O2-induced oxidative damage in rat intestinal epithelial IEC-6 cells. IEC-6 cells ...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-019-00354-x

    authors: Li J,Luo J,Zhang Y,Tang C,Wang J,Chen C

    更新日期:2020-02-01 00:00:00

  • Formation of multilayer rosettes in phytohemagglutinin-stimulated lymphocytes: correlation with chromatin condensation conformation in premature chromosome condensation.

    abstract::We found that the formation of multilayer rosettes by transformed human blood lymphocytes after phytohemagglutinin (PHA) stimulation is correlated with conformational changes of the chromatin as seen by premature chromosome condensation (PCC). The frequency distribution of grades of PCC and multilayer rosette formatio...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/BF00145027

    authors: Thomou H,Sambani C,Kitsiou P,Politis G

    更新日期:1988-07-01 00:00:00

  • On-line heat flux measurements improve the culture medium for the growth and productivity of genetically engineered CHO cells.

    abstract::With the increasingly competitive commercial production of target proteins by hybridoma and genetically engineered cells, there is an urgent requirement for biosensors to monitor and control on-line and in real time the growth of cultured cells. Since growth is accompanied by an enthalpy change, heat dissipation measu...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1023/A:1008038515285

    authors: Guan YH,Kemp RB

    更新日期:1999-07-01 00:00:00

  • Activated macrophages as a feeder layer for growth of resident cardiac progenitor cells.

    abstract::The adult heart contains a population of cardiac progenitor cells (CPCs). Growing and collecting an adequate number of CPCs demands complex culture media containing growth factors. Since activated macrophages secrete many growth factors, we investigated if activated isolated heart cells seeded on a feeder layer of act...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-014-9814-0

    authors: Sepúlveda DE,Cabeza Meckert P,Locatelli P,Olea FD,Pérez NG,Pinilla OA,Díaz RG,Crottogini A,Laguens RP

    更新日期:2016-08-01 00:00:00

  • The prophylactic use of antibiotics in cell culture.

    abstract::This article describes the historical development of the prophylactic use of antibiotics in cell culture as well as their effects on cells. The influence of antibiotics on cell morphology, cellular degeneration and cell death and cellular function is summarized. Cellular DNA as well as protein synthesis are affected w...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/BF00749764

    authors: Kuhlmann I

    更新日期:1995-06-01 00:00:00

  • Construction of recombinant monoclonal antibodies from a chicken hybridoma line secreting specific antibody.

    abstract::The chicken is a useful animal for the development of the specificantibodies against the mammalian conserved proteins. We generated twotypes of recombinant chicken monoclonal antibodies (mAbs), using a phagedisplay technique from a chicken hybridoma HUC2-13 which secreted themAb to the N-terminal of the mammalian prio...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1023/A:1008149815908

    authors: Nakamura N,Aoki Y,Horiuchi H,Furusawa S,Yamanaka HI,Kitamoto T,Matsuda H

    更新日期:2000-03-01 00:00:00

  • Evaluation of immunostimulatory effect of the arrowroot (Maranta arundinacea. L) in vitro and in vivo.

    abstract::Arrowroot (Maranta arundinacea. L) is an underutilized local crop potentially to be developed as carbohydrate source and functional food in Indonesia. The objectives of this research are to evaluate the immunostimulatory effects of arrowroot extracts in vitro by using animal cell culture techniques, and in vivo by usi...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-011-9403-4

    authors: Kumalasari ID,Harmayani E,Lestari LA,Raharjo S,Asmara W,Nishi K,Sugahara T

    更新日期:2012-03-01 00:00:00

  • Development of human monoclonal antibodies: A review.

    abstract::This article describes the current status in the development of human monoclonal antibodies. Over the last ten years a lot of information about the human immune system has emerged. Combining these with the many new (bio-)technologies it is plausible that the long awaited breakthrough of this technology is close. This ...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/BF00365341

    authors: Lindl T

    更新日期:1996-01-01 00:00:00

  • Productivity enhancement of recombinant protein in CHO cells via specific promoter activation by oncogenes.

    abstract::To construct a recombinant protein highly producing cell lines, we have previously developed the Oncogene Activated Production (OAP) system by using BHK-21 cells. Here we verified the availability of the OAP system in CHO cells. We firstly generated 'primed' ras amplified CHO cells, ras clone I, by introducing human c...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1023/A:1008048928053

    authors: Katakura Y,Seto P,Miura T,Ohashi H,Teruya K,Shirahata S

    更新日期:1999-09-01 00:00:00

  • Change in growth kinetics of hybridoma cells entrapped in collagen gel affected by alkaline supply.

    abstract::The growth yields for glucose and glutamine of murine hybridoma cells entrapped in collagen gel particles were examined during the growth phase. The immobilized hybridoma cells were cultivated in a fluidized bed fermenter where the medium was circulating to supply oxygen separately. Procedures to supply an alkaline so...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/BF00758178

    authors: Shirai Y,Yamaguchi M,Kobayashi A,Nishi A,Nakamura H,Murakami H

    更新日期:1994-01-01 00:00:00

  • Cytoglobin: a potential marker for adipogenic differentiation in preadipocytes in vitro.

    abstract::Obesity, mainly characterized by the excess fat storage, is a global health problem resulting in serious morbidity and mortality. Identification of molecular mechanisms in adipogenic differentiation pathway might lead to development of new strategies for diagnosis, prevention and therapy of obesity and associated dise...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-016-0047-2

    authors: Doğan A,Demirci S,Kıratlı B,Şahin F

    更新日期:2017-02-01 00:00:00

  • Highly passage of Spodoptera litura cell line causes its permissiveness to baculovirus infection.

    abstract::It is well known that the characteristics of cell lines possibly alter when cell lines are at high-passage number because of the environmental selection. We do not know whether non-permissive or low-permissive cell lines could become permissive or more permissive to virus infection after over-high passage. In the pres...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-008-9158-8

    authors: Zhang X,Lan W,Deng Y,Ma Y,Liu K,Peng J,Li Y,Hong H

    更新日期:2008-07-01 00:00:00

  • Cyanidin-3-glucoside enhances mitochondrial function and biogenesis in a human hepatocyte cell line.

    abstract::Mitochondrial dysfunction has been identified as one of the primary factors contributing to liver diseases. Pathways that control mitochondrial biogenesis are potential therapeutic targets for the amelioration of hepatocyte dysfunction and liver disease. Research on natural pharmacological agents that ameliorate liver...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-018-0242-4

    authors: Mogalli R,Matsukawa T,Shimomura O,Isoda H,Ohkohchi N

    更新日期:2018-12-01 00:00:00

  • Sericin in the isolating solution improves the yield of islets isolated from the pancreas.

    abstract::Approximately half of the transplantable pancreatic islet tissue is lost during isolation, including the digestion and purification steps. Modifying the isolation method could increase the yield. This would enable the one donor-one recipient concept and improve the therapeutic effects of islet transplantation. This st...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-016-9970-5

    authors: Yokoi S,Murakami M,Morikawa M,Goi T,Yamaguchi A,Terada S

    更新日期:2016-12-01 00:00:00

  • Small interfering RNA knockdown of mini-TyrRS and mini-TrpRS effects angiogenesis in human umbilical vein endothelial cells in hypoxic culture.

    abstract::Aim We studied the role of mini-TyrRS and mini-TrpRS in angiogenesis by using small interfering RNA-mediated mini-TyrRS/mini-TrpRS knockout in hypoxic culture of human umbilical vein endothelial cells. Methods SiRNA was used as the main method to inhibited the gene function. Silencing efficiency was assayed by real-ti...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-008-9151-2

    authors: Zeng R,Chen YC,Zeng Z,Liu R,Qiang O,Jiang XF,Liu XX,Li X,Wang HY

    更新日期:2008-03-01 00:00:00

  • Features of free software packages in flow cytometry: a comparison between four non-commercial software sources.

    abstract::Flow cytometers designed to analyze large particles are enabling new applications in biology. Data analysis is a critical component of the process FCM. In this article we compare features of four free software packages including WinMDI, Cyflogic, Flowing software, and Cytobank. ...

    journal_title:Cytotechnology

    pub_type: 杂志文章

    doi:10.1007/s10616-013-9609-8

    authors: Sahraneshin Samani F,Moore JK,Khosravani P,Ebrahimi M

    更新日期:2014-08-01 00:00:00