Interaction of Biomphalaria cells in primary cultures with Schistosoma mansoni sporocysts

eaaci2015

Culture of Intestinal Epithelial Cell Monolayers and Their Use in Multiplex Macromolecular Permeability Assays for In Vitro Analysis of Tight Junction Size Selectivity

 

Tight junctions form a selectively permeable barrier that limits paracellular flux across epithelial-lined surfaces. Small molecules (less than ∼8 Å diameter) can traverse the junction via the size- and charge-selective, high-conductance pore pathway. In contrast, the low-conductance leak pathway accommodates larger macromolecules (up to ∼100 Å diameter) and is not charge-selective. Flux across the tight junction-independent, high-conductance, non-selective, unrestricted pathway occurs at sites of epithelial damage.

 

Cytokines can regulate each of these pathways, but commonly used measures of barrier function cannot discriminate between tight junction regulation and epithelial damage. This article describes methods for culturing intestinal epithelial cell monolayers and assessing the impact of cytokine treatment on leak and unrestricted pathway permeabilities. © 2020 Wiley Periodicals LLC. Basic Protocol 1: Generation and culture of cell monolayers in Transwells Basic Protocol 2: Assessment of cytokine (IFNγ and TNF) treatment effects on barrier function Support Protocol: Immunofluorescent staining of monolayers Basic Protocol 3: Multiplex flux assay.

 

 

Interaction of Biomphalaria cells in primary cultures with Schistosoma mansoni sporocysts

Introduction: Biomphalaria snails may display varying levels of susceptibility to Schistosoma mansoni infection. We have been developing an in vitro model to study the interaction between the snail and the parasite, using tissue-derived cell cultures from Biomphalaria.
Methods: The digestive gland- and kidney-derived cells from primary cultures of resistant (B. tenagophila Taim) and susceptible (B. tenagophila HM and B. glabrata BH) strains of Biomphalaria were exposed to S. mansoni sporocysts.
Results: S. mansoni sporocysts were surrounded and encapsulated exclusively by cells derived from the digestive gland (DG) of B. tenagophila Taim. The process was followed by a marked decrease in the number of free sporocysts in the culture medium. The morphological characteristics of DG-derived cells in culture have been described.
Conclusions: Cells derived from DG (but not SK) primary cultures of B. tenagophila Taim may participate in S. mansoni sporocyst control.
eaaci2015

eaaci2015

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Comparison of VEGF-A secretion from tumor cells under cellular stresses in conventional monolayer culture and microfluidic three-dimensional spheroid models

 

Vascular endothelial growth factor (VEGF) is a major cytokine in tumor biology affecting tumor survival, aggressiveness and pro-angiogenetic activities. In addition, cellular stresses often result in aggressive pro-angiogenetic behavior in tumors. For in vitro study, conventional monolayer cell culture has been broadly exploited; however, it often provides limited information due to its different microenvironment from that in vivo. Recently, three-dimensional (3D) cell spheroid culture provides in vivo-like microenvironments to study tumor biology and their survival mechanisms with better predictive power.
In this work, vascular endothelial growth factor of type A (VEGF-A) secretion from osteosarcoma (MG-63) cells cultured using monolayer and 3D spheroid models under two stress conditions: nutrient deficiency (reduced serum culture) and hypoxia-inducible factor (HIF) inhibition (HIF inhibitor, YC-1) are characterized and systematically compared.
In order to obtain ample sample size for consistent characterization of cellular responses from cancer spheroids under the stresses and compare the responses to those from the conventional monolayer model, a microfluidic spheroid formation and culture device is utilized in the experiments.
In the analysis, cell viability is estimated from captured images, and quantification of VEGF-A secreted from the cells is achieved using enzyme-linked immunosorbent assay (ELISA). The experimental results show that the viabilities decrease when the cells face higher stress levels in both monolayer and 3D spheroid culture models; however, the VEGF-A secretion profiles between the cell culture models are different.
The VEGF-A secretion decreases when the cells face higher stress conditions in the monolayer cell culture. In contrast, for the 3D spheroid culture, the VEGF-A concentration decreases for low stress levels but increases while the stress level is high. The VEGF-A regulation in the 3D models mimics in vivo cases of tumor survival and can provide insightful information to investigate tumor angiogenesis in vitro. The approach developed in this paper provides an efficient method to quantitatively and statistically study tumor growth kinetics and stress responses from highly uniform samples and it can also be applied to compare the underlying biomolecular mechanisms in monolayer and 3D spheroid culture models to elucidate the effects of microenvironments on cellular response in cancer research.

Protection of human induced pluripotent stem cells against shear stress in suspension culture by Bingham plastic fluid

 

Suspension culture is an important method used in the industrial preparation of pluripotent stem cells (PSCs), for regenerative therapy and drug screening. Generally, a suspension culture requires agitation to keep PSC aggregates suspended and to promote mass transfer, but agitation also causes cell damage. In this study, we investigated the use of a Bingham plastic fluid, supplemented with a polysaccharide-based polymer, to preserve PSCs from cell damage in suspension culture. Rheometric analysis showed that the culture medium gained yield stress and became a Bingham plastic fluid, after supplementation with the polymer FP003.

 

A growth/death analysis revealed that 2 days of aggregate formation and 2 days of suspension in the Bingham plastic medium improved cell growth and prevented cell death. After the initial aggregation step, whereas strong agitation (120 rpm) of a conventional culture medium resulted in massive cell death, in the Bingham plastic fluid we obtained the same growth as the normal culture with optimal agitation (90 rpm). This indicates that Bingham plastic fluid protected cells from shear stress in suspension culture and could be used to enhance their robustness when developing a large-scale. This article is protected by copyright. All rights reserved.

 

Metabolic profiling of CHO cell cultures at different working volumes and agitation speeds using spin tube reactors

 

Culture systems based on spin tube reactors have been consolidated in the development of manufacturing processes based on Chinese hamster ovary (CHO) cells. Despite their widespread use, there is little information about the consequences of varying operational setting parameters on the culture performance of recombinant CHO cell lines. Here we investigated the effect of varying working volumes and agitation speeds on cell growth, protein production and cell metabolism of two clonally-derived CHO cell lines (expressing an IgG1 and a ‘difficult-to-express’ fusion protein). Interestingly, low culture volumes increased recombinant protein production and decreased cell growth, while high culture volumes had the opposite effect. Altering agitation speeds exacerbated or moderated the differences observed due to culture volume changes.

 

Combining low agitation rates with high culture volumes suppressed growth and recombinant protein production in CHO cells. Meanwhile, high agitation rates narrowed the differences in culture performance between low and high working volumes. These differences were also reflected in cell metabolism, where low culture volumes enhanced oxidative metabolism (linked to a productive phenotype) and high culture volume generated a metabolic profile that was predominately glycolytic (linked to a proliferative phenotype). Our findings indicate that the culture volume influence on metabolism modulates the balance between cell growth and protein production, a key feature that may be useful to adjust CHO cells towards a more productive phenotype.

 

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  • 5 g
  • Shipped within 1-2 weeks.

2-Amino-Isonicotinic Acid Ethyl Ester

20-abx188205
  • EUR 718.00
  • EUR 356.00
  • 100 g
  • 25 g
  • Shipped within 1-2 weeks.

2-Hydroxy-4-Amino Butanoic Acid

abx188257-1kg 1 kg
EUR 746
  • Shipped within 1-2 weeks.

4-Amino-3-phenylbutyric acid HCl

20-abx183511
  • EUR 230.00
  • EUR 732.00
  • EUR 356.00
  • 1 g
  • 25 g
  • 5 g
  • Shipped within 1-2 weeks.

Fmoc-21-Amino-4,7,10,13,16,19-Hexaoxaheneicosanoic Acid

20-abx184074
  • EUR 871.00
  • EUR 606.00
  • 1 g
  • 500 mg
  • Shipped within 1-2 weeks.

D-Amino-Acid Oxidase (DAO) Antibody

abx431198-200ul 200 ul
EUR 286
  • Shipped within 1-3 working days.

D-Amino-Acid Oxidase (DAO) Antibody

abx432587-200ul 200 ul
EUR 384
  • Shipped within 1-3 working days.

D-Amino-Acid Oxidase (DAO) Antibody

abx432588-200ul 200 ul
EUR 384
  • Shipped within 1-3 working days.

Human Amino Acid Metabolism Primer Library

HAMM-I 1 set
EUR 645

4-Amino-piperidine-4-carboxylic acid

F-3605.0001 1.0g
EUR 345
Description: Sum Formula: C6H12N2O2; CAS# [40951-39-1] net

4-Amino-piperidine-4-carboxylic acid

F-3605.0005 5.0g
EUR 1301
Description: Sum Formula: C6H12N2O2; CAS# [40951-39-1] net

4-Amino-piperidine-4-carboxylic acid

F-3605.0025 25.0g
EUR 5108
Description: Sum Formula: C6H12N2O2; CAS# [40951-39-1] net

(S)-(-)-2-Amino-4-pentenoic acid

GM0237-1G 1 g
EUR 90

(S)-(-)-2-Amino-4-pentenoic acid

GM0237-5G 5 g
EUR 237

Recombinant rat D-amino-acid oxidase

P1766 100ug Ask for price
  • Uniprot ID: O35078
  • Reconstitution: Metal affinity chromatography on Fn Super Capacity Column (Nickel)
Description: Recombinant protein for rat D-amino-acid oxidase

Native Porcine D-Amino Acid Oxidase

NATE-0180 400u
EUR 270

D-amino acid oxidase, human recombinant

P1052-10
EUR 164

D-amino acid oxidase, human recombinant

P1052-50
EUR 588

L-Amino Acid Assay Kit (Colorimetric)

MET-5054 100 assays
EUR 409

L-Amino Acid Assay Kit (Fluorometric)

MET-5055 100 assays
EUR 409

Branched Chain Amino Acid Assay Kit

MET-5056 192 assays
EUR 450

Anti-D-amino-acid oxidase antibody

STJ71455 100 µg
EUR 359

Anti-D Amino Acid Oxidase (2F5)

YF-MA12633 100 ug
EUR 363
Description: Mouse monoclonal to D Amino Acid Oxidase

Atpenin A5 (Synthetic)

2210-1000
EUR 457

Atpenin A5 (Synthetic)

2210-250
EUR 191

Synthetic pyrethroid insecticides

AT123 1mg
EUR 1368

Synthetic pyrethroid insecticide

AG123 1 mg
EUR 523

Synthetic Neurotensin (NT)

4-SPB203Hu01
  • EUR 368.80
  • EUR 202.00
  • EUR 1108.00
  • EUR 436.00
  • EUR 772.00
  • EUR 310.00
  • EUR 2620.00
  • 100 ug
  • 10ug
  • 1 mg
  • 200 ug
  • 500 ug
  • 50ug
  • 5 mg
  • Uniprot ID: P30990
  • Buffer composition: PBS, pH 7.4.
  • Form: Freeze-dried powder
  • Predicted Molecular Mass (KD): 1561.9Da
  • Isoelectric Point: 9.7
Description: Recombinant Human Neurotensin expressed in: E.coli

Synthetic Phytohemagglutinin (PHA)

4-SPB359Ge01
  • EUR 368.80
  • EUR 202.00
  • EUR 1108.00
  • EUR 436.00
  • EUR 772.00
  • EUR 310.00
  • EUR 2620.00
  • 100 ug
  • 10ug
  • 1 mg
  • 200 ug
  • 500 ug
  • 50ug
  • 5 mg
  • Uniprot ID: Inquire
  • Buffer composition: PBS, pH 7.4, containing 0.01% SKL, 1mM DTT, 5% Trehalose and Proclin300.
  • Form: Freeze-dried powder
  • Predicted Molecular Mass (KD): Inquire
  • Isoelectric Point: Inquire
Description: Recombinant Pan-species Phytohemagglutinin expressed in: E.coli

Synthetic Triacylated Lipopeptide

VAdv-Ly0036 1 mg
EUR 1628
Description: Synthetic triacylated lipopeptide, a synthetic lipopeptide and a TLR2/1 agonist vaccine adjuvant.

D-amino acid oxidase activator Polyclonal Antibody

42143-100ul 100ul
EUR 333

Human D-amino acid oxidase activator (DAOA)

1-CSB-EP006495HU
  • EUR 380.00
  • EUR 214.00
  • EUR 1309.00
  • EUR 560.00
  • EUR 873.00
  • EUR 262.00
  • 100ug
  • 10ug
  • 1MG
  • 200ug
  • 500ug
  • 50ug
  • MW: 45.1 kDa
  • Buffer composition: Tris-based buffer with 50% glycerol.
Description: Recombinant Human D-amino acid oxidase activator(DAOA) expressed in E.coli

Human xcitatory amino acid transporter 3 (SLC1A1)

1-CSB-EP021432HU
  • EUR 380.00
  • EUR 214.00
  • EUR 1309.00
  • EUR 560.00
  • EUR 873.00
  • EUR 262.00
  • 100ug
  • 10ug
  • 1MG
  • 200ug
  • 500ug
  • 50ug
  • MW: 37.5 kDa
  • Buffer composition: Tris-based buffer with 50% glycerol.
Description: Recombinant Human xcitatory amino acid transporter 3(SLC1A1),partial expressed in E.coli

Mouse Excitatory amino acid transporter 2 (Slc1a2)

1-CSB-EP021433MO
  • EUR 505.00
  • EUR 265.00
  • EUR 1827.00
  • EUR 766.00
  • EUR 1218.00
  • EUR 335.00
  • 100ug
  • 10ug
  • 1MG
  • 200ug
  • 500ug
  • 50ug
  • MW: 37.6 kDa
  • Buffer composition: Tris-based buffer with 50% glycerol.
Description: Recombinant Mouse Excitatory amino acid transporter 2(Slc1a2),partial expressed in E.coli