Combining TruCulture® and OptiMAP to Profile Human Immunity

Shen-An Hwang, Dominic Eisinger, and Samuel LaBrie


TruCulture® is a whole blood collection and culture system designed for use at clinical sites. TruCulture outperforms traditional peripheral blood mononuclear cell (PBMC) culturing methods, with improved reproducibility and higher levels of stimulation (Duffy et. al. Clinical Immunology 2017). The TruCulture proprietary culture medium ensures minimal non-specific immune activation, delivering a steady baseline and decreased variation across study populations. OptiMAP is a multiplex immunoassay panel that measures 13 cytokines of the major immune response pathways (TH1, TH2, TH17, and monocyte/macrophage). Using TruCulture and OptiMAP, we analyzed healthy donor whole blood responses to common stimulants: T cell activators, TLR agonists, and superantigens. The subjects ranged from 24 to 70 years old and consisted of 48% males and 52% females. The results indicated that healthy donors responded similarly to stimulation with anti-CD3/CD28 and staph enterotoxin type B (SEB) in production of T cell cytokines (IFN-γ, IL-2, and IL-17). The response to SEB is positively correlated to the monocyte population in whole blood. T cell responses to SEB and CytoStim (an antibody based alternative to SEB) are similar in production of IFN-γ and IL-17, but CytoStim had a reduced stimulation of IL-2. As expected, all healthy donors responded to a TLR-4 agonist (LPS) by producing high levels of TNF-α, IL-6, and IL-1β. LPS stimulated production of IFN-γ was similar to SEB, most likely due to bystander activation. The TLR-7 agonist (gardiquimod, GDQ) stimulated IL-10 levels at a level comparable to LPS. These data illustrate the utility of combining TruCulture and OptiMAP to investigate human immune stimulation pathways for clinical research.

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How TruCulture Works

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.

TruCulture Procedure, Collect


Draw 1 mL of blood directly into the TruCulture Tube and break off the plunger.

TruCulture Procedure, MIX

02. MIX

Gently invert tube to mix 3 to5 times

TruCulture Procedure, Incubate


Place tube in 37ºC heat block for up to 24 or 48 hours.

TruCulture Procedure, separate


Manually insert valve to separate supernatant from the cells. Collect supernatant and cell layer for downstream analysis.


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

PBMCs vs. TruCulture

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.

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 PBMCs

  • 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.
PBMC vs TruCulture whole blood collection and stimulation

Learn why TruCulture Whole Blood Culture System is a more relevant model of human leukocyte function than the traditional method of cultured PBMCs.