General Process-Based Model for Describing the Metabolic Shift in Microbial Cell Cultures

November 12, 2020
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eaaci2015

Cell Cultures for Virology: Usability, Advantages, and Prospects

Virus detection in natural and clinical samples is a complicated problem in research and diagnostics. There are different approaches for virus isolation and identification, including PCR, CRISPR/Cas technology, NGS, immunoassays, and cell-based assays. Following the development of genetic engineering methods, approaches that utilize cell cultures have become useful and informative. Molecular biology methods allow increases in the sensitivity and specificity of cell cultures for certain viruses and can be used to generate reporter cell lines.
These cell lines express specific reporter proteins (e.g., GFP, luciferase, and CAT) in response to virus infection that can be detected in a laboratory setting. The development of genome editing and synthetic biology methods has given rise to new perspectives regarding the design of virus reporter systems in cell cultures. This review is aimed at describing both virology methods in general and examples of the development of cell-based methods that exist today.

A General Process-Based Model for Describing the Metabolic Shift in Microbial Cell Cultures

The metabolic shift between respiration and fermentation at high glucose concentration is a widespread phenomenon in microbial world, and it is relevant for the biotechnological exploitation of microbial cell factories, affecting the achievement of high-cell-densities in bioreactors. Starting from a model already developed for the yeast Saccharomyces cerevisiae, based on the System Dynamics approach, a general process-based model for two prokaryotic species of biotechnological interest, such as Escherichia coli and Bacillus subtilis, is proposed. The model is based on the main assumption that glycolytic intermediates act as central catabolic hub regulating the shift between respiratory and fermentative pathways. Furthermore, the description of a mixed fermentation with secondary by-products, characteristic of bacterial metabolism, is explicitly considered.
The model also represents the inhibitory effect on growth and metabolism of self-produced toxic compounds relevant in assessing the late phases of high-cell density culture. Model simulations reproduced data from experiments reported in the literature with different strains of non-recombinant and recombinant E. coli and B. subtilis cultured in both batch and fed-batch reactors. The proposed model, based on simple biological assumptions, is able to describe the main dynamics of two microbial species of relevant biotechnological interest. It demonstrates that a reductionist System Dynamics approach to formulate simplified macro-kinetic models can provide a robust representation of cell growth and accumulation in the medium of fermentation by-products.

Developmentally-Inspired Biomimetic Culture Models to Produce Functional Islet-Like Cells From Pluripotent Precursors

Insulin-producing beta cells sourced from pluripotent stem cells hold great potential as a virtually unlimited cell source to treat diabetes. Directed pancreatic differentiation protocols aim to mimic various stimuli present during embryonic development through sequential changes of in vitro culture conditions. This is commonly accomplished by the timed addition of soluble signaling factors, in conjunction with cell-handling steps such as the formation of 3D cell aggregates. Interestingly, when stem cells at the pancreatic progenitor stage are transplanted, they form functional insulin-producing cells, suggesting that in vivo microenvironmental cues promote beta cell specification. Among these cues, biophysical stimuli have only recently emerged in the context of optimizing pancreatic differentiation protocols.
This review focuses on studies of cell-microenvironment interactions and their impact on differentiating pancreatic cells when considering cell signaling, cell-cell and cell-ECM interactions. We highlight the development of in vitro cell culture models that allow systematic studies of pancreatic cell mechanobiology in response to extracellular matrix proteins, biomechanical effects, soluble factor modulation of biomechanics, substrate stiffness, fluid flow and topography. Finally, we explore how these new mechanical insights could lead to novel pancreatic differentiation protocols that improve efficiency, maturity, and throughput.

Effects of FGFR inhibitors TKI258, BGJ398 and AZD4547 on breast cancer cells in 2D, 3D and tissue explant cultures

Purpose: Fibroblast growth factor receptors (FGFR) and pathways are important players in breast cancer (BC) development. They are commonly altered, and BCs exhibiting FGFR gene amplification are currently being studied for drug development. Here, we aimed to compare the effects of three FGFR inhibitors (FGFRis), i.e., non-selective TKI258 and selective BGJ398 and AZD4547, on different BC-derived cell lines (BCCs) and primary tissues.
Methods: The human BCCs MCF-7 and MDA-MB-231(SA) (wild-type FGFR) and MFM223 (amplified FGFR1 and FGFR2) were analyzed for FGFR expression using qRT-PCR, and the effects of FGFRis on FGFR signaling by Western blotting. The effects of FGFRis on proliferation, viability, migration and invasion of BCCs were assessed in 2D cultures using live-cell imaging, and in 3D cultures using phenotypic analysis of organoids. To study radio-sensitization, FGFRi treatment was combined with irradiation. Patient-derived BC samples were treated with FGFRis in explant cultures and immunostained for Ki67 and cleaved caspase 3.
Results: We found that all FGFRis tested decreased the growth and viability of BC cells in 2D and 3D cultures. BGJ398 and AZD4547 were found to be potent at low concentrations in FGFR-amplified MFM233 cells, whereas higher concentrations were required in non-amplified MCF7 and MDA-MB-231(SA) cells. TKI258 inhibited the migration and invasion, whereas BGJ398 and AZD4547 only inhibited the invasion of MDA-MB-231(SA) cells. FGFRi treatment of MCF7 and MFM223 cells enhanced the inhibitory effect of radiotherapy, but this effect was not observed in MDA-MB-231(SA) cells. FGFRi-treated primary BC explants with moderate FGFR levels showed a tendency towards decreased proliferation and increased apoptosis.
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ASE-9234 1 vial (1 x 10^6)
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cDNA from Human Adult Normal Tissue: Peripheral Blood Leukocyte
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Description: Can be used for various studies in the realm of gene expression, both normal and pathological. It is an excellent control and suitable for educational purposes.

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Total Protein from Human Adult Normal Tissue: Peripheral Blood Leukocyte
P1234148 1 mg
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Membrane Protein from Human Adult Normal Tissue: Peripheral Blood Leukocyte
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Conclusions: Our results indicate that, besides targeting FGFR-amplified BCs with selective FGFRis, also BCs without FGFR amplification/activation may benefit from FGFRi-treatment. Combination with other treatment modalities, such as radiotherapy, may allow the use of FGFRis at relatively low concentrations and, thereby, contribute to better BC treatment outcomes.

Inflammatory lncRNA AK039862 regulates paraquat-inhibited proliferation and migration of microglial and neuronal cells through the Pafah1b1/Foxa1 pathway in co-culture environments

 

Emerging evidences having suggested that particular lncRNAs have a potential effect on PD progression through provoking damage and inflammatory responses of microglia/ dopaminergic cells. In addition, paraquat can be accumulated in human body through various approaches and have an increased risk for Parkinson’s disease. However, the specific role and mechanism of lncRNA related to neurotoxic in the progression of PD is unclear. In our study, a mouse PD model was established induced by the intraperitoneal injection of paraquat (5 mg/kg and 10 mg/kg) every three days (10 times).
We determined differential expression of lncRNA AK039862 and its potential targeted genes Pafah1b1/Foxa1 in PD mouse model, then we used fluorescence in situ hybridization (FISH) to visualize the cellular distribution of AK039862. Short interfering RNAs (siRNAs) and overexpression plasmids were designed for knockdown or overexpression of AK039862. To simulate the coexisting dopaminergic cells and microglia cells in vitro, we applied several non-contact co-culture models, including conditioned medium and Transwell co-culture systems. Cytotoxicity of PQ was evaluated using bv2 cells with the concentrations: 30, 60 μM, and mn9d cells with the concentrations: 50, 100 μM. As a result, we depicted multiple interesting individual and interactive features of inflammatory lncRNA AK039862 involved in PQ-induced cellular functional effects.
First, we detected that AK039862 contributed to the neuronal injury process in PQ-treated mice and co-localization of AK039862 with dopaminergic cells in vivo. And interestingly, we demonstrated that PQ significantly inhibited microglia and dopaminergic cells proliferation and microglia migration in vitro. Further research indicated that the PQ-induced low expression of AK039862 rescued microglia proliferation and migration inhibition via the AK039862/Pafah1b1/Foxa1 pathway.
Meanwhile, AK039862 also participated in the interaction between microglia and dopaminergic cells with PQ treatment in non-contact co-culture models. In summary, we found that PQ inhibited the proliferation and migration of microglial cells, and elucidated AK039862 played a key role in PQ-induced neuroinflammatory damage through Pafah1b1/Foxa1. Finally, inflammatory AK039862 is involved in the complex communication between microglia and dopaminergic cells in the environment of PQ damage.

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EUR 416.75
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EUR 401
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Human Umbilical Vein Endothelial Cells
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Human B-lymphocyte antigen CD19 (CD19)
1-CSB-CF004888HU
  • EUR 1298.00
  • EUR 682.00
  • EUR 798.00
  • 1MG
  • 200ug
  • 500ug
Description: Recombinant Human B-lymphocyte antigen CD19(CD19),partial expressed in in vitro E.coli expression system

Human B-lymphocyte antigen CD19 (CD19)
1-CSB-CF004888HUc7
  • EUR 965.00
  • EUR 665.00
  • EUR 715.00
  • 1MG
  • 200ug
  • 500ug
Description: Recombinant Human B-lymphocyte antigen CD19(CD19),partial expressed in in vitro E.coli expression system

human umbilical vein endothelial cells, Peptide Aptamer, unlabeled
AP-316-U 5 mg Ask for price

Sheep Red Blood Cells
20R-RS001 10 ml
EUR 521
Description: Human IgG Senstitized Gluteraldehyde Stabilized Sheep Red Bllood Cells

Bovine Red Blood Cells
88R-B002 20 ml
EUR 273
Description: Glutaraldehyde-stabilized freshly prepared Bovine Red Blood Cells

Chicken Red Blood Cells
88R-C002 20 ml
EUR 273
Description: Glutaraldehyde-stabilized freshly prepared Chicken Red Blood Cells

Dog Red Blood Cells
88R-D004 10 ml
EUR 273
Description: Glutaraldehyde-stabilized freshly prepared Dog Red Blood Cells

Goat Red Blood Cells
88R-G003 20 ml
EUR 273
Description: Glutaraldehyde-stabilized freshly prepared Goat Red Blood Cells

Hamster Red Blood Cells
88R-H001 5 ml
EUR 273
Description: Glutaraldehyde-stabilized freshly prepared Hamster Red Blood Cells

Horse Red Blood Cells
88R-H002 20 ml
EUR 273
Description: Glutaraldehyde-stabilized freshly prepared Horse Red Blood Cells

Hamster Red Blood Cells
88R-H003 10 ml
EUR 489
Description: Washed and freshly prepared 10% suspension of Hamster Red Blood Cells

Mouse Red Blood Cells
88R-M001 10 ml
EUR 295
Description: Washed and freshly prepared 10% suspension of Mouse Red Blood Cells

Monkey Red Blood Cells
88R-M003 5 x 2 ml
EUR 273
Description: Glutaraldehyde-stabilized freshly prepared Monkey Red Blood Cells

Mouse Red Blood Cells
88R-M004 5 x 2 ml
EUR 273
Description: Glutaraldehyde-stabilized freshly prepared Mouse Red Blood Cells

Pig Red Blood Cells
88R-P002 20 ml
EUR 273
Description: Glutaraldehyde-stabilized freshly prepared Pig Red Blood Cells

Rabbit Red Blood Cells
88R-R001 20 ml
EUR 273
Description: Glutaraldehyde-stabilized freshly prepared Rabbit Red Blood Cells

Rat Red Blood Cells
88R-R002 5 x 2.0ml
EUR 273
Description: Glutaraldehyde-stabilized freshly prepared Rat Red Blood Cells

Rat Red Blood Cells
88R-R005 25 ml
EUR 327
Description: Washed and freshly prepared 10% suspension of Rat Red Blood Cells

Sheep Red Blood Cells
88R-S001 100 ml
EUR 512
Description: Washed and freshly prepared 10% suspension of Sheep Red Blood Cells

Sheep Red Blood Cells
88R-S003 20 ml
EUR 273
Description: Glutaraldehyde-stabilized freshly prepared Sheep Red Blood Cells

Sheep Red Blood Cells
88R-S004 10 ml
EUR 521
Description: Glutaraldehyde-stabilized freshly prepared albumin sensitised Sheep Red Blood Cells

Sheep Red Blood Cells
88R-S005 10 ml
EUR 446
Description: Glutaraldehyde-stabilized freshly prepared tanned Sheep Red Blood Cells

Turkey Red Blood Cells
88R-T001 20 ml
EUR 273
Description: Glutaraldehyde-stabilized freshly prepared Turkey Red Blood Cells

Mouse Monoclonal Anti-Human CD19 FITC/CD2 PE
CD19-D1-50 50 tests
EUR 651

Mouse Monoclonal Anti-Human CD19 FITC/CD3 PE
CD19-D2-50 50 tests
EUR 651

Mouse Monoclonal Anti-Human CD19 FITC/CD5 PE
CD19-D3-50 50 tests
EUR 651

Unprocessed Human Cord Blood from IRB-approved donors, Fresh
FC-001-F10 - Ask for price

Unprocessed Human Cord Blood from IRB-approved donors, Fresh
FC-001-F100 - Ask for price

Unprocessed Human Cord Blood from IRB-approved donors, Fresh
FC-001-F50 - Ask for price

Unprocessed Human Cord Blood from IRB-approved donors, Fresh
FC-001-F80 - Ask for price

Mouse Umbilical Cord FibrOut: Fibroblast Inhibitory System (for 500 ml medium)
7-15184 1 Kit Ask for price

Rat Umbilical Cord FibrOut: Fibroblast Inhibitory System (for 500 ml medium)
7-15264 1 Kit Ask for price

human umbilical vein endothelial cells, Peptide Aptamer, FITC labelled
AP-316-F 1 mg Ask for price

CORNING®HUMAN UMBILICAL VEIN ENDOTHELIAL CELLS, HUVEC-2 CELLS (>5X105 CELLS), 1 CRYOVIAL/PACK
354151 1/pk
EUR 388
Description: Permeable Support - DL; Coated Transwells - DL

B-Lymphocyte Antigen CD19 (CD19) Antibody
abx011636-100ul 100 ul
EUR 411

B-Lymphocyte Antigen CD19 (CD19) Antibody
abx159376-100ul 100 ul
EUR 356

B-Lymphocyte Antigen CD19 (CD19) Antibody
abx159377-100ul 100 ul
EUR 356

B-Lymphocyte Antigen CD19 (CD19) Antibody
abx159378-100ul 100 ul
EUR 356

B-Lymphocyte Antigen CD19 (CD19) Antibody
abx159379-100ul 100 ul
EUR 356

B-Lymphocyte Antigen CD19 (CD19) Antibody
abx159380-100ul 100 ul
EUR 439

B-Lymphocyte Antigen CD19 (CD19) Antibody
abx159381-100ul 100 ul
EUR 356

Rabbit Red Blood Cells (100%)
20-abx800015
  • EUR 648.00
  • EUR 509.00
  • 100 ml
  • 50 ml

Sheep Red Blood Cells (100%)
20-abx800032
  • EUR 356.00
  • EUR 314.00
  • 100 ml
  • 50 ml
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