Use of the mitochondria toxicity assay for quantifying the viable cell density of microencapsulated jurkat cells

2013 ◽  
Vol 29 (4) ◽  
pp. 986-993 ◽  
Author(s):  
M. Werner ◽  
K. Biss ◽  
V. Jérôme ◽  
F. Hilbrig ◽  
R. Freitag ◽  
...  
Keyword(s):  
Cell Density ◽  
Viable Cell ◽  
Viable Cell Density ◽  
Jurkat Cells ◽  
Toxicity Assay ◽  
Mitochondria Toxicity

scholarly journals Mapping the molecular basis for growth related phenotypes in industrial producer CHO cell lines using differential proteomic analysis

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Laura Bryan ◽  
Michael Henry ◽  
Ronan M. Kelly ◽  
Christopher C. Frye ◽  
Matthew D. Osborne ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Density ◽  
Cell Lines ◽  
Viable Cell ◽  
Viable Cell Density ◽  
High Peak ◽  
Proteomic Profiling ◽  
Cho Cell ◽  
Extended Culture ◽  
Key Pathways

Abstract Background The ability to achieve high peak viable cell density earlier in CHO cell culture and maintain an extended cell viability throughout the production process is highly desirable to increase recombinant protein yields, reduce host cell impurities for downstream processing and reduce the cost of goods. In this study we implemented label-free LC-MS/MS proteomic profiling of IgG4 producing CHO cell lines throughout the duration of the cell culture to identify differentially expressed (DE) proteins and intracellular pathways associated with the high peak viable cell density (VCD) and extended culture VCD phenotypes. Results We identified key pathways in DNA replication, mitotic cell cycle and evasion of p53 mediated apoptosis in high peak VCD clonally derived cell lines (CDCLs). ER to Golgi vesicle mediated transport was found to be highly expressed in extended culture VCD CDCLs while networks involving endocytosis and oxidative stress response were significantly downregulated. Conclusion This investigation highlights key pathways for targeted engineering to generate desirable CHO cell phenotypes for biotherapeutic production.

scholarly journals Directional Selection of Microbial Community Reduces Propionate Accumulation in Glycerol and Glucose Anaerobic Bioconversion Under Elevated pCO2

2021 ◽  
Vol 12 ◽  
Author(s):  
Pamela Ceron-Chafla ◽  
Yu-ting Chang ◽  
Korneel Rabaey ◽  
Jules B. van Lier ◽  
Ralph E. F. Lindeboom
Keyword(s):  
Microbial Community ◽  
Cell Viability ◽  
Cell Density ◽  
Viable Cell ◽  
Viable Cell Density ◽  
Directional Selection ◽  
Adaptive Laboratory Evolution ◽  
Glycerol Conversion ◽  
Microbial Groups ◽  
Selection Of

Volatile fatty acid accumulation is a sign of digester perturbation. Previous work showed the thermodynamic limitations of hydrogen and CO2 in syntrophic propionate oxidation under elevated partial pressure of CO2 (pCO2). Here we study the effect of directional selection under increasing substrate load as a strategy to restructure the microbial community and induce cross-protection mechanisms to improve glucose and glycerol conversion performance under elevated pCO2. After an adaptive laboratory evolution (ALE) process, viable cell density increased and predominant microbial groups were modified: an increase in Methanosaeta and syntrophic propionate oxidizing bacteria (SPOB) associated with the Smithella genus was found with glycerol as the substrate. A modest increase in SPOB along with a shift in the predominance of Methanobacterium toward Methanosaeta was observed with glucose as the substrate. The evolved inoculum showed affected diversity within archaeal spp. under 5 bar initial pCO2; however, higher CH4 yield resulted from enhanced propionate conversion linked to the community shifts and biomass adaptation during the ALE process. Moreover, the evolved inoculum attained increased cell viability with glucose and a marginal decrease with glycerol as the substrate. Results showed differences in terms of carbon flux distribution using the evolved inoculum under elevated pCO2: glucose conversion resulted in a higher cell density and viability, whereas glycerol conversion led to higher propionate production whose enabled conversion reflected in increased CH4 yield. Our results highlight that limited propionate conversion at elevated pCO2 resulted from decreased cell viability and low abundance of syntrophic partners. This limitation can be mitigated by promoting alternative and more resilient SPOB and building up biomass adaptation to environmental conditions via directional selection of microbial community.

scholarly journals Effect of temperature and agitation rate variation on the growth of NS0 cell line in monoclonal antibody production

2019 ◽  
Author(s):  
Shamitha Shetty
Keyword(s):  
Cell Density ◽  
Cell Metabolism ◽  
Viable Cell ◽  
Viable Cell Density ◽  
Effect Of Temperature ◽  
Research Report ◽  
Rate Variation ◽  
Agitation Rate ◽  
Shake Flasks ◽  
Culture Growth

The quality and yield of the monoclonal antibodies produced in a cGMP environment is heavily influenced by the bioprocess-related parameters which impact the cell growth and metabolism of the mammalian cell cultures. This research report describes a study conducted to examine the effects of varying temperature and RPM set points on viable cell density and viability of NS0 cultures. All cultures were grown in 250 mL shake flasks (working vol. 100 mL). To separately analyze the effects of temperature and agitation rate on NS0 cell metabolism, flask stage cultures were evaluated in triplicates at two cultivation temperatures (36 °C and 38 °C) and two agitation rates (120 RPM and 160 RPM) while controls were maintained at 37 °C and 140 RPM for both the conditions using an incubator. Flasks were sampled every 24 h and analyzed for viable cell density and % viability. Additional data was collected on pH, pO2, pCO2, osmolality, glucose, lactate, glutamate and glutamine levels in the culture. It was observed that variations in temperature has the greatest effect on viable cell density and viability and varying agitation rates had minimal effect on growth of cultures. A temperature set point of 38 °C is detrimental to the culture growth. The control set points proved to be optimal for this process.

E2F-1 overexpression increases viable cell density in batch cultures of Chinese hamster ovary cells

2008 ◽  
Vol 138 (3-4) ◽  
pp. 103-106 ◽  
Author(s):  
Brian S. Majors ◽  
Nilou Arden ◽  
George A. Oyler ◽  
Gisela G. Chiang ◽  
Nels E. Pederson ◽  
...  
Keyword(s):  
Cell Density ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster Ovary Cells ◽  
Chinese Hamster ◽  
Viable Cell ◽  
Viable Cell Density ◽  
Ovary Cells ◽  
Batch Cultures

scholarly journals FACTORS AFFECTING CHINESE HAMSTER OVARY CELL PROLIFERATION AND VIABILITY

2019 ◽  
Vol 1 ◽  
pp. 245
Author(s):  
Alina Rekena ◽  
Dora Livkisa ◽  
Dagnija Loca
Keyword(s):  
Cell Proliferation ◽  
Cell Line ◽  
Cell Density ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster ◽  
Viable Cell ◽  
Viable Cell Density ◽  
Fed Batch ◽  
Cho Cell ◽  
Decline Phase

Advantageous cultivation procedures for the Chinese hamster ovary (CHO) cells are necessary for the productive commercial production of biopharmaceuticals. A main challenge that needs to be addressed during the process development is the differences in each cell line requirements concerning the nutrients and feed strategies in order to achieve the desired growth characteristics. Therefore, within the current research, a naïve high cell density serum free suspension adapted CHO cell line was tested with glucose and glutamine rich feeds in fed-batch Erlenmeyer shake flask cultures. Glucose consumption rate was adjusted to develop the optimal feed strategies. Obtained results indicated that high glucose and l-glutamine feeding did not improve maximum viable cell density compared to the control samples. During the exponential phase, cell proliferation and viability of all feeds showed no statistically significant difference. Instead, the fed-batch processes tested led to statistically significant differences in viable cell density and cell viability during the decline phase, compared to control (batch) culture. The difference between glucose and glutamine feeding was indistinguishable, most probably due to the concentration imbalance with the rest of the nutrients in feed. The overall study presented a method to slow down the decrease in CHO cell proliferation and viability during the decline phase, instead of increasing the maximum cell density at the plateau phase. 

Analysis of Stored Osteochondral Allografts at the Time of Surgical Implantation

2005 ◽  
Vol 33 (10) ◽  
pp. 1479-1484 ◽  
Author(s):  
R. Todd Allen ◽  
Catherine M. Robertson ◽  
Andrew T. Pennock ◽  
William D. Bugbee ◽  
Frederick L. Harwood ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Cell Viability ◽  
Cell Density ◽  
Tissue Bank ◽  
Viable Cell ◽  
Viable Cell Density ◽  
Proteoglycan Synthesis ◽  
Superficial Zone ◽  
Chondrocyte Viability ◽  
Osteochondral Allografts

Background To date, the morphological, biochemical, and biomechanical characteristics of articular cartilage in osteochondral allografts that have been stored have not been fully described. Hypothesis Osteochondral allografts procured and stored commercially for a standard period as determined by tissue banking protocol will have compromised chondrocyte viability but preserved extracellular matrix quality. Study Design Controlled laboratory study. Methods Unused cartilage from 16 consecutive osteochondral allografts was sampled during surgery after tissue bank processing and storage. Ten grafts were examined for cell viability and viable cell density using confocal microscopy, proteoglycan synthesis via 35SO4 uptake, and glycosaminoglycan content and compared with fresh cadaveric articular cartilage. Biomechanical assessment was performed on the 6 remaining grafts by measuring the indentation stiffness of the cartilage. Results The mean storage time for the transplanted specimens was 20.3 ± 2.9 days. Chondrocyte viability, viable cell density, and 35SO4 uptake were significantly lower in allografts at implantation when compared to fresh, unstored controls, whereas matrix characteristics, specifically glycosaminoglycan content and biomechanical measures, were unchanged. In addition, chondrocyte viability in the stored allografts was preferentially decreased in the superficial zone of cartilage. Conclusion Human osteochondral allografts stored for a standard period (approximately 3 weeks) before implantation undergo decreases in cell viability, especially in the critically important superficial zone, as well as in cell density and metabolic activity, whereas matrix and biomechanical characteristics appear conserved. The exact clinical significance of these findings, however, is unknown, as there are no prospective studies examining clinical outcomes using grafts stored for extended periods. Clinical Relevance Surgeons who perform this procedure should understand the cartilage characteristics of the graft after 21 days of commercial storage in serum-free media.

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