Creative Biolabs combines the discovery of synthetic gene circuits for cancer immunotherapy with the development of immune response, establishing novel therapies for tumors with strong and specific immune responses. The deep expertise is the foundation of our collaboration with leaders in the global pharmaceutical industry. We are committed to providing the power of in vitro Th17 differentiation assay to open up new avenues in the fight against cancer.

Introduction to Th17 Differentiation

In general, helper T cells have been divided into Th1 and Th2 cells according to their specific cytokine expression profiles. Recently, a new helper T cell (Th17) that can produce IL-17 has also been identified. Studies have shown that Th17 cells are not only involved in host defense, but can also induce the development of several inflammatory diseases, autoimmune diseases, and tumors. As a result, it is particularly important to identify the factors responsible for human Th17 cell differentiation.

Fig.1 Th17 Cells Stimulate IL-6 Production by Bronchial Epithelial Cells.¹

At Creative Biolabs, we are exploring a completely new in vitro Th17 differentiation assay to characterize helper T cell properties and to analyze the differentiation and function of Th17 cells. Typically, we differentiate CD4+ T cells in vitro and then establish a range of techniques, including but not limited to, real-time quantitative PCR, ELISA, and immunofluorescence analysis of gene expression. Finally, a co-culture analysis system will be generated to evaluate TH cell subset functions by monitoring primary bronchial epithelial cell activities.

Main Procedures

• Naïve CD4+ T Cell Culture and Differentiation

Density gradients were used to isolate mononuclear cells from cord blood samples. CD45RA+ T cells were stimulated with soluble anti-CD3 and anti-CD28 antibodies. TH17 cells differentiate with the help of various cytokines and antibodies, such as IL-6, IL-1b, anti-IL-12 antibodies, and anti-IFN-γ antibodies. The best dose volume for each cytokine and antibody will be determined in advance by using our serial dose-titration assays. Naïve CD4+ T cells are cultured in optimal density under different conditions, usually using serum-free medium or RPMI 1640 medium in our labs.

• Cytokine Production Detection

In Creative Biolabs, the content of supernatant cytokines is normally determined by ELISA. Results are quantified by measuring the absorbance value at 450 nm.

• Real-Time Quantitative RT-PCR Analysis

RNA is extracted from cultured CD4+ T cells and cDNA is synthesized for real-time quantitative PCR analysis. These specific primers for IL-17, IL-22, IL-23R, RORC2, and IFN-γ will be added to the PCR system for amplification. We used relative quantitative methods to process the data.

Advantages of In Vitro Th17 Differentiation Assay Service at Creative Biolabs

• A positive feedback loop

The core competencies at Creative Biolabs are focused on developing, performing, and analyzing in vitro Th17 differentiation. We often design, develop, and run tests that produce high throughput readings.

• A full-scale service panel

We support a wide range of in vitro Th17 differentiation services, ranging from off-the-shelf to fully customized development projects. Just tell us your specific requirements and we will send you the results as soon as possible.

• Cell source diversity

We can utilize a wide range of cell types in our in vitro Th17 differentiation assay services, including primary, PBMC, and patient-derived cells.

Creative Biolabs is your trusted CRO partner. Our dedicated inspection development team will use novel technologies and proprietary in vitro Th17 differentiation analysis solutions to overcome any obstacles and build reliable, repeatable inspection assays based on your needs. Please feel free to contact us. Our technical team will work closely with you to ensure a smooth project execution and help you achieve your research goals.

Reference

1. Burgler, S.; et al. Differentiation and functional analysis of human TH17 cells. Journal of Allergy and Clinical Immunology, 2009, 123(3): 588-595.

For Research Use Only | Not For Clinical Use