TruCulture
A Whole Blood Collection and Culture Tube for Standardized Functional Immunophenotyping Procedures
TruCulture tubes are designed to capture immune cell activity at the time and place of sample collection, thereby minimizing the bias and variability introduced by sample shipping and manipulation. These revolutionary tubes consist of an integrated whole-blood collection and leukocyte culture system that is reliable, simple to use and does not require specialized laboratory equipment. TruCulture enables researchers to reproducibly ascertain information about immune responses to stress or stimulation, including responses to novel therapeutics. The ex vivo TruCulture procedure preserves physiological cellular interactions to more accurately reflect the complexities of the human immune system, bringing added value to immune monitoring in clinical trials.
TruCulture Procedure
TruCulture tubes are pre-loaded with cell culture media and immune stimulant(s) or drug candidates. Blood is drawn directly into the TruCulture tube and incubated in a dry heat block. Supernatants are collected by simply inserting a valve separator to separate cells from the culture supernatant.
01. COLLECT
Draw 1 mL of blood directly into the TruCulture Tube and break off the plunger.
02. MIX
Gently invert tube to mix 3 to 5 times
03. INCUBATE
Place tube in 37ºC heat block for up to 24 or 48 hours.
04. SEPARATE
Manually insert valve to separate supernatant from the cells. Collect supernatant and cell layer for downstream analysis.
TruCulture Tube Components
TruCulture Applications
TruCulture has been utilized as a whole-blood stimulation system by researchers and drug developers in several fields to reliably measure immune response for the following applications.
- Pharmacodynamics (including dose response)
- Functional immune cell analysis
- Disease characterization
- Patient stratification
- Genotype – to – Phenotype association studies
- Vaccine Development
- Antigen recall
- Early safety studies
- Profile the functional immune status of clinical trial participants
Our partners at, The Institut Pasteur, demonstrated the indispensable benefits of TruCulture in The Milieu Intérieur (“Environment Within”) project. Briefly, the Milieu Intérieur project is working to establish the determinants of a healthy immune response by identifying the genetic, epigenetic and environmental factors that contribute to the observed heterogeneity of immune responses. The challenge of the project is to establish the boundaries of a healthy immune response, and to that end, the Milieu Intérieur Project recruited 1,000 healthy individuals stratified across age (five-decades of life from age 20 – 69), and gender (1:1 gender ratio). TruCulture tubes were an integral part of the Milieu Intérieur clinical study, used as a standardized method for monitoring human response to immune stimulation.
To define the boundaries of a healthy immune response, The Milieu Intérieur project combined the use of TruCulture whole blood culture system with the use of stimuli to define the protein signatures induced by (i) medically relevant bacteria, fungi and viruses; (ii) agonists specific for defined host sensors; (iii) clinically employed cytokines; and (iv) activators of T cell immunity.
View an Overview of The Milieu Intérieur Project:
Vaccine Applications Include
- Antigen recall
- Early safety studies
- Profile the functional immune status of clinical trial participants
The use of TruCulture for vaccine development has been highlighted by two key examples:
- Whole‐blood cytokine secretion assay as a high‐throughput alternative for assessing the cell‐mediated immunity profile after two doses of an adjuvanted SARS‐CoV‐2 recombinant protein vaccine candidate. (2022) De Rosa SC, et al. Clinical & Translational Immunology. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8752373/
- Safety and immunogenicity of SARS-CoV-2 recombinant protein vaccine formulations in healthy adults: interim results of a randomized, placebo-controlled, phase 1-2, dose-ranging study. (2021) Goepfert PA, et al. Lancet Infectious Disease. https://doi.org/10.1016/S1473-3099(21)00147-X
Supplementary Material including TruCulture details and figures, view here.
For the two-dose cohort, pre-vaccination and post-vaccination blood samples (D1, D22, D36) were collected into three types of TruCulture tubes:
1) Recombinant SARS-CoV-2 spike protein
2) Negative control (TruCulture media only)
3) Positive control (SEB + CD28)
Cytokines from supernatants were measured at Rules-Based Medicine. Production of Th1 associated cytokines (IFNg, IL-2 and TNFa) were elevated compared to Th2 associated cytokines (IL-4, IL-5, IL-13) on D22 and D36 whereas neutralizing antibodies were not significantly detected until D36. The AS03 adjuvant formulation generated stronger humoral and cell mediated immunity signals relative to the AF03 adjuvant. This study illustrates that TruCulture can be a vital tool for vaccine clinical trials to profile antigen specific T cell responses.
2. Flu Antigen Recall Responses using TruCulture with OptiMAP and SIMOA Analysis(2019) Hwang SA, et al.
The recall response of flu hemagglutinin (HA) antigens was examined in healthy adults vaccinated with the 2018-2019 annual flu vaccine. The TruCulture tubes used were:
- Null (TruCulture media only)
- Staphylococcal enterotoxin B (SEB)
- Null tubes spiked with 1.25mg each of 4 different recombinant HA proteins.
All samples were incubated for 48 hours at 37oC. Supernatant were collected and analyzed using the OptiMAP panel. No inflammatory cytokines were detected in the null tubes from either the baseline or 2 weeks post-vaccination samples. There were no differences in cytokine production in the SEB stimulated samples between the baseline and 2 weeks post-flu vaccination timelines. For flu HA stimulated samples, there was a significant increase in production of GM-CSF, IFN-, IL-1‑, IL-6, IL-12p70, IL-23, and TNF-a at 2 weeks post-flu vaccination compared to samples collected at baseline. Additionally, flu HA stimulated samples post vaccination also demonstrated increased production of type I inteferons, as analyzed by SIMOA. These data demonstrate that TruCulture in conjunction with OptiMAP and SIMOA are useful tools for investigating antigen specific responses.
Advantages of TruCulture
- Integrated closed sterile instant whole blood collection and culture system.
- Standardized to ensure consistent performance across multiple users and clinical sites.
- Reliable, easy to use, and reproducible – eliminates the need for cell manipulation.
- Retains all blood components, granulocytes, platelets, red blood cells, soluble factors and Fc receptor expressing cells.
- Only inexpensive heat block needed, no lab equipment nor centrifugation steps.
- Has been successfully deployed in hundreds of clinical drug trials.
Disadvantages of Traditional PBMC
- Separate blood collection and specialized cell culture procedures.
- Extensive manipulation, processing, and often freezing/shipping prior to culturing.
- Requires technical expertise with increased variability across users and clinical sites.
- Requires CO2 incubator, biosafety cabinet, centrifuge, media, and cell culture plastics.
- Culture procedures/conditions are difficult to standardize for clinical trial applications.
- Open, less sterile, artificial system.
- Poor reproducibility.
Traditional pharmacodynamic whole blood experiments are generally short in duration (2-6 hrs) as a consequence of poor culture conditions leading to premature termination of the normal physiological immune response. A longer more robust response usually provides higher sensitivity and greater relevance than short incubation times that may only lead to the release of stored pre-synthesized mediators.
View Peer-Reviewed Publications Citing TruCulture
Bibliography
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Milieu Intérieur: Personalized Immune Response Monitoring
“We hope to advance the promise of personalized medicine with functional biomarkers.”
Dr. Matthew Albert, Director of the Department of Immunology; Co-Director and Founder of the Center for Human Immunology at Institut Pasteur; Adjunct Faculty at Cochin Hospital
Applications of TruCulture in Chronic infection
“What makes our project unique is our ability to induce standardized immune responses…possible because of the reproducibility of the TruCulture system.”
Dr. Darragh Duffy, Laboratory of Dendritic Cell Biology, Department of Immunology, Institut Pasteur
Statistical Analysis and Mathematical Modeling of Data Derived from TruCulture
“[Using TruCulture] we get high quality data with very little variation.”
Pr. Magnus Fontes, Director of the International Group for Data Analysis, Institut Pasteur
Stimulants
Estimated lead time for TruCulture Tube Delivery is 6-8 Weeks from receipt of PO, signed contract where applicable, and Shipping Form (2 weeks for LPS and Null Tubes which are routinely kept in stock)
Part # | Stimulus | Description | Type | Final Concentration (After 1mL of blood is added) |
---|---|---|---|---|
782-001277 | Interferon beta (IFN-beta) | Type I interferon, modulator of for example T lymphocyte responses | Cytokine | 1000 IU/mL |
782-001278 | Interleukin-1beta (IL-1beta) + tumor necrosis factor- alpha (TNF-alpha) | 2 synergistically acting pro-inflammatory cytokines (weak to modest immune cell activation) | Cytokine | IL-1beta: 25 ng/mL TNF-alpha: 10 ng/mL |
782-001295 | TNF-alpha | Pro-inflammatory cytokine; weak activator of mediator synthesis when used alone | Cytokine | 10 ng/mL |
782-001086 | Null | Pure (proprietary) TruCulture media without stimulants | Negative Control | n/a |
782-001291 | NegCo | TruCulture media without stimulants, specially formulated for premium and custom tubes | Negative Control including custom excipients | 0.033 % HSA |
782-001272 | Adenosine Triphosphate (ATP) + Lipopolysaccharide (LPS-EB) | ATP modulates via purinergic receptors (such as P2X7) LPS-induced activation of cells of the innate part of the immune system | NLRP3 Inflammasome / TLR4 Ligand | ATP: 2.5 mM LPS-EB: 10 ng/mL |
782-001273 | Lauroyl-γ-D-glutamyl-meso-diaminopimelic acid (C12-iE-DAP) | Dipeptide representing bacterial peptido-glycan, activator of NOD1 (intracellular pattern recognition receptor) | NOD Ligand | 4 µg/mL |
782-001259 | Zymosan | ß-glucan particles (fractions of yeast cell walls); triggers phagocytosis via TLR2, Dectin, and CD11/18 | Phagocyte activator | 5 µg/mL |
782-001275 | HKEB | Heat killed preparation of the gram negative bacterium, E. Coli O111:B4. Triggers phagocytosis via TLR2, TLR4, and various others | Phagocyte activator | 10^7 cells/mL |
782-001125 | anti-CD3 + anti-CD28 | Two antibodies triggering T-cell activation via the signaling unit of the T-cell receptor complex (CD3) + co-activation (intensifying T-cell responses, adding activities of Th2 and Treg) via CD28 | T-Cell activation | CD3: 0.2 µg/mL CD28: 0.33 µg/mL |
782-001202 | anti-CD3 | T-cell activation via the signaling unit of the T-cell receptor complex (CD3) | T-Cell activation | 0.2 µg/mL |
782-001416 | TStim | TCR-MHC cross-linking Mabs plus co-stim | T-Cell activation | Stim: 1.5 µL/mL coStim: 0.33 µg/mL |
782-001274 | Fibroblast-Stimulating Lipopeptide (FSL-1) |
Synthetic analogue of microbial lipoprotein; agonist of TLR2/TLR6 |
TLR2 Ligand | 2 µg/mL |
782-001282 | Polyinosinic : polycytidylic acid (Poly I:C) | Analogue of double-stranded RNA, mimics the presence of viral infection. Activator of TLR3 | TLR3 Ligand | 20 µg/mL |
782-001087 | Lipopolysaccharide (LPS) | Bacterial endotoxin (E.coli, O55:B5) that elicits a strong innate immune response | TLR4 Ligand | 0.1 µg/mL |
782-001261 | LPS-EB | Bacterial endotoxin (E.coli, O111:B4) that elicits a strong innate immune response | TLR4 Ligand | 10 ng/mL |
782-001455 | LPS+Tstim | Bacterial edotoxin (E.coli, O55:B5) that elicits a strong innate response in addition to a T cell response by crosslinking the T cell receptor and MHC on monocytes/macrophages | TLR4 Ligand + T cell receptor ligand |
LPS: 0.1 µg/mL Stim 1.5 µL/mL |
782-001269 | Gardiquimod | Synthetic agonist of TLR7 (responding to single-stranded RNA, for example) | TLR7 Ligand | 3 µM |
782-001264 | Resiquimod (R848) | Synthetic agonist of TLR7 and TLR8 (both responding to single-stranded RNA) | TLR7/8 Ligand | 1 µM |
If the stimulant you’d like to use in our TruCulture tubes is not listed, please fill out this form to request it. One of our experts will contact you to discuss the feasibility of your request. Thank you for your interest in TruCulture.
RBM provides quantitative measurement of immune related proteins from TruCulture supernatants. Whether your specific application requires a broad range or a focused set of analytes, RBM’s panels are designed to meet your needs.
TruCulture® OptiMAP is an immunoassay testing panel validated and optimized to quantify the effect of TruCulture stimulations. It is a cost-effective test comprised of 13 biomarkers that captures the diversity of responses to microbial, viral, and T cell stimulants.
OptiMAP Biomarkers
MAP | Volume Required |
---|---|
OptiMAP v. 1.0 | TruCulture Supernatant 100 μL |
OptiMAP Design
A total of 106 immune-related biomarkers were tested on TruCulture samples taken from healthy subjects. A careful selection of the key stimulants listed below were used to monitor a wide range of immune response pathways. From the 106 candidate biomarkers, we selected a subset of 13 assays that were useful in distinguishing responses to individual stimuli while also revealing population variances.
Microbial stimulants: LPS, IL-1β, TNF-α, BCG, HK C. albicans, HK E. coli, HK S. aureus
Viral stimulants: Poly IC, R848, IFN- α/β
T cell stimulants: SEB, anti-CD3/CD28
For a more comprehensive understanding of functional immune cell activity, TruCulture supernatant can be tested on any of our Human MAP products. The tubes are also compatible with flow cytometry and gene expression analysis.
Multiplex immunoassay analysis of TruCulture supernatants is conducted in our CLIA-certified testing laboratory.
Every order starts with a quote.
To get your quote, contact RBM or choose an option below:
Co-Developing new tubes
To co-develop Truculture tubes of interest, please contact us.
RBM can evaluate your stimuli of interest for sterility, solubility, dose response, short and long-term stability, and reproducibility.
Support
RBM is proud to provide excellent customer support
Let us know how we can help you using one of the following options:
Already have a quote number? Great!
Application Notes
Measuring Antigen Recall to SARS-CoV-2 Spike Protein with TruCulture
TruCulture, TruCulture Webinars, Webinars
Protocols for Processing TruCulture Samples at Clinical Sites
Application Notes, TruCulture
Gene Expression Analysis of RBM’s TruCulture Samples with the NanoString nCounter Panel
Application Notes, TruCulture
RNA Extraction/Purification Protocol for TruCulture Samples App Note
Application Notes, TruCulture
Vaccine Development with TruCulture Application Note
Application Notes, TruCulture
Standard Flow Cytometry Protocol for Analysis of Surface Markers for TruCulture
Application Notes, TruCulture
TruCulture Webinars
Measuring Antigen Recall to SARS-CoV-2 Spike Protein with TruCulture
TruCulture, TruCulture Webinars, Webinars
Investigating Immune Response with TruCulture & NanoString nCounter
TruCulture, TruCulture Webinars, Webinars
A Reproducible Immunomonitoring Method for Multi-Center Clinical Studies
TruCulture Webinars, Webinars
Standardized Assays for Personalized Immune Response Monitoring
TruCulture, TruCulture Webinars, Webinars