Next Generation Cell Recovery for Immuno-Oncology
Scientists from Charles River Laboratories and Curiox Biosystems collaborated to characterize the performance and workflow of Laminar Wash™ systems in isolating and preparing tumor infiltrating leukocytes (TILs) from anti-PD1-challenged CT26 mice, as a model system.
Compared to conventional centrifuge-based methods, Laminar Wash technology:
- Improved recovery of viable TILs from dissociated tumors
- Reduced inter-operator variation in TIL recovery
- Shortened labor-intensive hands-on washing steps
The novel Laminar Wash system represents an elegant, reproducible, and partially automated approach to recovering viable and well-defined TIL subpopulations for quantification and analysis by multicolor flow cytometry.
Developing a 43 Color Panel on Cytek Aurora Using Laminar Wash for Sample Prep
Successfully executing a 43 color panel is a technically demanding task. One that is made more difficult when there are confounding issues like non-specific binding and cell debris.
The methods described in this new publication in Cytometry Part A show that the preparation of the samples was very similar to traditional cell staining with the exception that Laminar Wash Technology was used to prepare the cells in addition to the large number of antibody types used. Because the results that were presented showed incredibly high reproducibility, including at low cell numbers, it would seem safe to posit that preparing cells with Laminar Wash technology is key in providing quality reproducible data.
Innovative Tools for Reliable, Reproducible and Efficient Gene Silencing Experiments
siPOOLs™: Complex siRNA Pools for Specific and Reliable RNA Interference
siPOOLs are defined, high complexity pools of 30 siRNAs designed for maximal transcript coverage.
Avoid off-target effects
Efficient knockdown (at low nM)
Efficient siRNA transfection of primary cells and cell lines
SilenceMag siRNA Delivery Reagent is based on the Magnetofection™ technology. Simple, rapid and easy to use, SilenceMag is serum compatible and non toxic. Specifically designed for siRNA transfection, SilenceMag gives reliable high protein knockdown at very low doses of siRNA in numerous cell types (primary cells, hard-to-transfect & cell lines).
- High protein knockdown efficiency
- For all siRNA applications
- No off-target effects
- Serum compatible & non toxic
- Simple, rapid and easy-to-use
Lullaby is the ideal siRNA transfection reagent for gene silencing. Relying on the TEE-technology, it has been successfully tested on numerous cell lines, reaching up to 90% gene silencing with high reproducibility and a very low toxicity. RNA interference is a powerful technique to shut down genes expression in cells and organisms.
Improved Lentiviral Transduction of Stem Cells
ViroMag Stem Transduction Enhancer
ViroMag Stem, a Magnetofection-based Lentiviral Transduction Enhancer, enables improved viral driven genetic modification in a wide range of stem cells in different applications, such as ex vivo gene therapy and cell therapy.
ViroMag Stem, as a stabilized magnetic nanoparticles formulation, offers a simple and reproducible method for increasing lentiviral infection and transduction of difficult cell types such as CD34+ hematopoietic stem cells, both cell lines and primary cells, in cell culture plate using the Magnetofection technology.
This method combines maximum cell viability and high transduction efficiency while cell phenotype and differentiation potential are not affected.
- Increase lentiviral transduction efficiency
- Concentrate viral dose, promote and accelerate the transduction process
- Efficient for hard-to-infect and non-permissive cells
- Synchronise cells adsorption/infection
ViroMag Stem outperforms lentiviral mediated transduction efficiency in CD34+ cell lines
KG1a CD34+ cells were infected at MOI 5 (5 viral particles per cell) using a HIV-SFFV-GFP lentivirus in presence of viral enhancers at the recommend doses. 72 hours after transduction, % of GFP positive cells (A) and total intensity (B) were analyzed by flow cytometry.
Laminar Wash™ Centrifuge-Free Wash of Cells and Nuclei
Faster and automatible, Laminar Wash™ systems deliver higher quality data through better cell retention, better preservation of cellular physiology and viability, and improved study-to-study and operator-to-operator reproducibility.
mRNAs Optimized for Improved Stability and Performance
We now offer mRNAs that mimic fully processed mature mRNAs.
These mRNAs are stabilized with 5’ Cap 1 structure and 3’ poly(A) tail and are optimized to yield improved stability and performance.
They are modified with 5-methoxyuridine (5moU replaces U) to reduce innate immune responses. We also offer unmodified mRNAs.
Reporter gene mRNAs: ideal controls to study transfection efficiency
Genome editing mRNA: Cas9 mRNA for CRISPR Genome editing
Vaccine mRNAs: ideal as controls for immunization or vaccine studies
Lullaby Stem siRNA Transfection Reagent
Lullaby Stem siRNA Transfection Reagent is ideal for gene silencing in Stem Cells. Lullaby Stem enables siRNA and miRNA transfection of multipotent and embryonic stem cells with high efficiency and very low toxicity. Its lipid-specific formulation protects small RNA from extracellular degradation, transports them across cell membranes and efficiently releases them into stem cell cytoplasm according to the TEE-technology.
Non-viral Vaccine Carrier Adjuvants for DNA or Protein Delivery
The new VaxOZ vaccine adjuvants from OZ Bioscience are perfect for any immunization experiment.
Currently, adjuvants remain to be the most important strategy to improve the efficiency of conventional and next-generation vaccines. Genetic adjuvant is an immunological agent uses in combination with a specific antigen in order to boost the immune response. The direct consequence is to produce more antibodies and longer-lasting immunity than with the antigen alone and thus minimizing the dose of antigen needed to the vaccine. In particular, DOTAP (1,2-dioleoyl-3-trimethylammonium-propane) based cationic lipids induce maturation of dentritic cells, which then traffic to the draining lymph nodes to activate antigen-specific T cells.
In vivo grade manufactured and quality controlled, the VaxOZ adjuvants are intended for research purposes only.
CaLiVax DOTAP cationic lipid-based composition for liposome-mediated DNA or protein vaccines.
PolyVax-CPO Vaccine Adjuvant is a cationic polymer genetic adjuvant that associates with plasmid DNA to form an efficient polymer-based nanoparticle delivery system (NPD).
CNE-CPO (Cationic Nano Emulsion) Vaccine Adjuvant is an oil-in-water Cationic Nano Emulsion made of squalene droplets and cationic polymers in a continuous aqueous phase. CNE-CPO Vaccine Adjuvant is biodegradable, an important advantage over alternative oils that have been used in emulsion adjuvants.
Enhancing Cell or Nuclei Recovery in Single Cell Multi-omics
Single-cell genomics and proteogenomics workflows can be greatly improved by the Laminar Wash™ technology.
The Laminar Mini is the perfect tool for single-cell sequencing experiments. It washes 8 samples simultaneously in a few minutes. It replaces centrifugation and preserves precious samples at low cell numbers. The workflow is easier and faster.
In the recent webinar, Curiox, Biolegend and ImmunAI presented data on increased cell retention compared to centrifugation, higher signal-to-noise ratios and time savings.
Automatic Live Cell Imaging System: Celloger Mini
Celloger Mini is a compact and easy-to-use live cell imaging system.
Compact design and easily fits in CO2 incubators
Positioning multiple area up to 96 well
Time-lapse image capturing & video clip
Convenient user software to operate the system
Providing the thermo-hygrometer data in real time
Automatic, Accurate and User-Friendly Cell Counter FACSCOPE B
FACSCOPE B has unique propoerties for rare cell populations and does not require any manual focusing.
Accurate and fast; choose between three different modes: Quick, Normal and Precise
Rare populations: count rare cells in the precise mode, allowing up to 3,6 µl volume
Convenient to use: automatic focusing
Enhance Lentiviral Transduction in Stem- and Immune Cells using LentiBlast Premium
The Lentiblast Premium Transduction Enhancer is designed to enhance viral transduction in many types of primary cells – adherent and suspension, especially also in CD34+ hematopoietic stem cells or T-lymphocytes. The image shows data on the effect of using LentiBlast Premium in CD34+ stem cells at a low viral particle/cell number ratio.
LentiBlast Premium is a patented reagent from OZ Biosciences with the following key benefits:
Higher efficiency in hard-to-transduct cells
High viability; non-toxic reagent
Save effort and money; use lower viral titers
Easy to use protocol
Magnetofection™ for Enhanced Transfection Efficiency in Primary and Hard-to-Transfect Cells
Magnetic Assisted Transfection is a novel technology developed by OZ Biosciences. It has a proven high efficiency in delivering molecules into challenging cells.
Magnetic nanoparticles are associated with nucleic acids (naked or pre-complexed with a transfectionreagent or viruses) by salt-induced aggregation and electrostatic interactions.
Magnetic force drives these complexes towards the target cells, allowing a rapid concentration of the vector dose onto cells
The cellular uptake of the genetic materials is accomplished by endocytosis and pinocytosis
Nucleic acids are released in the cytoplasm by flip-flop mechanism or proton sponge effect*
* Plank et al , Adv. Drug Deliv. Rev. (2011), 63(14-15):1300-31
New Transfection and Transduction Reagents with Unique Technologies
The reagents from OZ Biosciences covers a large range of applications:
In vivo Delivery
Laminar Wash Sample Preparation for Single-Cell Sequencing
Single-cell genomics is rapidly growing. Nature Methods named multimodal omics the method of the year for 2019. And as more scientists use single-cell techniques, they find that sample preparation is very important to achieve high-quality data.
Single cell sequencing can suffer from poor data quality due to clumping of cells, incomplete washing of cells leaving cell-free nucleic acids, and poor retention of low numbers of cells. Washing via centrifugation causes stress on the cells in samples and can cause erroneous gene expression results.
Sample prep is extremely important in single-cell genomics, especially for single cell gene expression and immune profiling. In this area, mRNA transcripts are being barcoded and cells can experience mRNA leakage (causing increased background) or mRNA degradation (causing less signal). Unlike with flow cytometry, in single-cell sequencing it is not possible to gate out debris or groups of cells. The cell preparation must be as clean as possible and consist of single cells.
By using the Laminar Wash system scientists can see time savings and less manual manipulation.
Laminar Wash systems can wash cells without putting stress on cells that may impact gene expression, cell integrity and viability.
Nuclei Isolation Using Laminar Wash
PBMCs were lysed in bulk in tube with Lysis Buffer, quenched with Wash Buffer and centrifuged
Cells are further either washed twiced in tube by centrifuge, OR plated in Laminar Wash plate to settle for 20min, washed 7x in a Laminar Wash HT1000 System with PBS + 0.04% BSA
Reliable, Automated and Reproducible Flow Cytometry
The unique Laminar Wash™ technology simplifies and automates flow cytometry workflows for CAR-T-cell manufacturing. Data improves reproducibility, saves time and gives tighter populations. Laminar Wash™ replaces centrifugation-based stain&wash and is more gentle to cells. Time saving in this workflow was 50 min/assay.
CAR-T cell therapy biotech company AdiCet Bio improved data by automating of a part of their flow cytometry assay. They saw improved consistency and are planning full automation using a Laminar Wash™ system.
Change the way you process cells to get reliably consistent data in flow cytometry
The DA-Cell Laminar wash technology replaces centrifugation by using a novel DA-Cell technology, which is gentle to cells, giving higher retention.
- A wide range of cell numbers, from single-cell to 10 million cells per well
- No stress on cells from centrifugation and improved viability
- Reduced debris and aggregation of cells
- Cleaner and faster washing
Enhanced viability and retention
siRNA Off-Target Effects; Causes, Extent & Impact
siPOOLs™ are innovative, complex siRNA pools which reduce false positives & false negatives in siRNA knockdown experiments
Individual siRNAs or low complexity siRNA pools containing 3-4 siRNAs often hit multiple off-target genes and exhibit variable target gene knock-down. Short interfering RNA (siRNA) pools, siPOOLs are highly complex and defined pools of 30 siRNAs, each siRNA present at picomolar working concentrations.
dilutes the off-target signature of each siRNA, increasing on-target specificity
ensures co-operative knock-down of the target gene, producing more robust and reliable results.
Download the new TechNote:
Off-Target Effects; Causes, Extent & Impact
Hannus et al NAR