Introduction to γδT Cells

The reason γδT cells have garnered widespread attention lies in their core characteristic of not requiring HLA antigen presentation.

This trait enables them to excel in reducing the risk of graft-versus-host disease (GVHD), significantly enhancing the overall safety of therapies and opening up broader application prospects for the 

treatment of various diseases. Compared with traditional TCR-based T cell therapies, γδT cells can recognize and target a variety of tumor antigens in an MHC-independent manner,

and they demonstrate unique advantages particularly in the treatment of solid tumors. Additionally, combining γδT cells with CAR-T technology can further strengthen their tumor-targeting capability,

paving a more promising path for cancer immunotherapy.

Introduction to Huachen Bio hyperClone® Human γδT Cell 
Activation/Expansion Cocktail Kit

In the field of immune cell therapy, γδT cells are emerging as a key research direction for the next generation of immune cell therapies due to their characteristic of not requiring HLA antigen presentation.
As a leading enterprise providing solutions in the cell therapy field, HUACHEN Bio has developed two advanced clinical-grade methods for the efficient activation and expansion of γδT cells, which are based on the novaT-36 medium and pureSep-γδT purification reagent. These methods offer an innovative full-process solution for the clinical application in this field.

hyperClone Human γδT Activation/Expansion Cocktail Kit

Using fresh/frozen PBMCs or CBMCs as the starting cells, purified γδT cells are easily obtained by efficiently and rapidly depleting αβT cells from whole blood using the pureSep-γδT Purification Reagent (αβT Depletion Reagent). Subsequently, the zoledronic acid activation method (1st Generation) and anti-CD3/CD28 activation method (2nd Generation) are applied to obtain γδT cells with high proliferation, high purity, and strong cytotoxicity. The two expansion methods provided by HUACHEN Bio each have their own focuses, offering flexible options for different research needs.

1st Generation γδT Cell Kit: Zoledronic Acid Activation Method

No pre-purification is required. Zoledronic acid is added during the activation process to expand Vδ2 cells and some Vδ1 cells, with the total number of cells achievable to expand 100-300 times.

The cell quantity can be scaled up by increasing the culture volume. On day 14 (D14), the cell purity exceeds 90% with high cytotoxic activity.

The 1st Generation Kit is suitable for both fresh and cryopreserved cells.

Photos of γδT Cell Culture

Cell Culture Data

Flow Cytometry Assay

2nd Generation γδT Cell Kit: Antibody Activation Protoco

The pureSep-γδT purification reagent efficiently and rapidly removes αβT cells from whole blood.

It achieves high-purity γδT cell enrichment based on the principle of density gradient negative selection, laying a foundation for ultra-high-purity expansion of γδT cells.

In subsequent steps, an antibody activation protocol is adopted to expand both Vδ2 cells and a subset of Vδ1 cells while maintaining high purity and functional integrity.

The entire process uses a pure cytokine formulation without introducing chemical reagent stimulation.

After 7 days (d7) of culture, the purity exceeds 90%, and after 14 days (d14), the purity exceeds 98%,

making it highly suitable for large-scale clinical applications of allogeneic γδT cells.

The enrichment of γδT cells prior to culture ensures stable expansion efficiency across different donors,

effectively addressing the common issue of donor individual variability in primary cell expansion and providing strong support for the reproducibility of research.

The second-generation kit, with the integration of the pureSep-γδT purification reagent, is suitable for use with fresh peripheral blood or umbilical cord blood.

The novaT-36 medium is a key support for these two expansion methods. Specially formulated, it is serum-free, xeno-free, and has a defined chemical composition, making it a preferred option for clinical-grade applications.

This medium can support the growth of both Vδ2 and Vδ1 cell subsets simultaneously, ensuring high purity and functional integrity of the expanded cells.

Photograph of γδT Cell Culture

Cell Culture Data

Day 7 Flow CytometryResults

Day 14 Flow CytometryResults

γδT cells were cultured until Day 14 (D14), then co-incubated 
with K562 cells for 20 hours to conduct a tumor cytotoxicity assay.