Serum-free Gamma Delta T Cells (γδT cell) Induction and Expansion Medium Kit
A new GMP medium specifically designed for Gamma Delta T cells
Used to induce and expand fresh or frozen human peripheral blood mononuclear cells into γδT cells in vitro efficiently and conveniently.
Description
| ltem No. | Product Name | Application | Storage conditions |
| NC0102 | Serum-free medium for NK cells | γδT cell culture in vitro | 2~8’C |
| AN0110 | γδT cell induction and expansion kit | YC00T: Add at the start of culture | -20’C |
| YC005: Add to culture medium for use |
Product Advantages
- It is used to induce and expand fresh and frozen human peripheral blood mononuclear cells into γδT cells in vitro. After 14-16 days of culture, PBMC can achieve more than 200-fold cell expansion and more than 85% positive rate.
- Pure factor induction, no trophoblast cells, simple and easy culture method, basically uniform cell state, according to the recommended process of rehydration, no need for cell counting.
- With the newly upgraded NK5.0 basal culture medium, the continuous expansion ability of cells is greatly improved.
Product Performance
1. Cell culture morphology
Day 3
Day 5
Day 7
Day 9
Day 11
Day 14
2. Case presentation
Sample 1: Fresh peripheral blood mononuclear cell sample from a 25-year-old healthy male. The sample was inoculated according to the total number of 4E7 cells, cultured for 14 days, and 9 billion cells were harvested, with a positive rate of 90.01%.
Sample 1: Fresh peripheral blood
Sample 2: A 35-year-old healthy male cryopreserved peripheral blood mononuclear cell sample was inoculated according to the total number of 4E7 cells and cultured for 14 days. 8.2 billion cells were harvested, with a positive rate of 90.29%.
Sample 2: Cryopreserved peripheral blood
Sample 3: A frozen peripheral blood mononuclear cell sample from a 27-year-old healthy male was inoculated according to the total number of 4E7 cells, cultured for 14 days, and 7 billion cells were harvested, with a positive rate of 83.51%.
Sample 3: Cryopreserved peripheral blood
Sample 4: Fresh peripheral blood mononuclear cell sample from a 38-year-old healthy male. It was inoculated according to the total number of 4E7 cells, cultured for 14 days, and 8.1 billion cells were harvested, with a positive rate of 92.66%.
Sample 4: Fresh peripheral blood
Sample 5: Fresh peripheral blood mononuclear cell sample from a 27-year-old healthy male was inoculated according to the total number of 6E7 cells, cultured for 14 days, and 8.36 billion cells were harvested, with a positive rate of 89.98%.
Sample 5: Fresh peripheral blood
γδT Cells: The "Special Forces" of T Cell Subsets – A Rising Star in Immunotherapy
"Special Forces" of the immune system
As a unique subset of T cells, γδT cells have gained a reputation as the “special forces” of the immune system. Their ability to recognize antigens in an MHC-unrestricted manner sets them apart, making them a powerful and versatile tool in the field of immunotherapy. With their potential to offer new therapeutic strategies for cancer and other diseases, γδT cells have attracted growing attention from research institutions and pharmaceutical companies alike. According to market forecasts, the global γδT cell therapy market is expected to exceed USD 5 billion by 2025, with China emerging as the second-largest market, growing at a compound annual rate of 28%.
However, a major hurdle to the commercialization of γδT therapies lies in the difficulty of efficient in vitro expansion. In response to this challenge, YOCON has developed a γδT Cell Expansion Culture Kit, powered by our newly upgraded NK5.0 basal medium. This system significantly enhances the sustained expansion potential of γδT cells. Within 14–16 days of culture, PBMCs can achieve over 200-fold expansion with a γδT cell purity exceeding 85%. This breakthrough addresses the core issues of low expansion efficiency and purity, providing a robust foundation for large-scale manufacturing and clinical application of γδT cell therapies.
1. What is the purity of γδT cells after 20 days of culture? Is there a big difference in killing efficiency compared to 14 days?
For cord blood-derived γδT cells, maturity is typically achieved around day 20, with purity reaching approximately 85% and cytotoxicity over 90% at an effector-to-target (E:T) ratio of 40:1. Peripheral blood-derived γδT cells usually mature by day 14–15, with a decline in phenotype starting around day 18, potentially reducing purity by about 10%.
2. When is the logarithmic growth phase of γδT cells during culture?
For peripheral blood-derived γδT cells, the exponential growth phase generally occurs between days 5 and 7. A significant growth surge is often observed following the medium replenishment on day 7.
3. Is an initial seeding density of 2×10⁶/mL mandatory?
The recommended seeding density is within the range of 2–2.5×10⁶ cells/mL. Deviations—either too high or too low—can negatively affect both the final yield and the proportion of γδT cells.
4. At what time point should cytotoxicity assays be conducted?
Short-term killing assays are typically conducted at 4–6 hours, while long-term effects are evaluated at 18–24 hours. Given the mechanism of γδT cells, an incubation of 8–20 hours is often recommended to better reflect their cytotoxic potential.
5. Can cord blood be used for γδT cell culture?
Yes, cord blood is a viable source for γδT cell culture. However, due to its limited availability, peripheral blood is generally preferred for routine expansion.
6. Which surface markers should be analyzed during γδT cell preparation?
Flow cytometry is typically used to assess CD3+ and γδTCR+ (specifically Vδ2+) populations.
7. How important is the freshness of the blood sample? Can frozen PBMCs be used? What if there’s no autologous plasma?
Fresh samples generally yield better culture outcomes. However, frozen PBMCs can also be effectively used, provided they have been cryopreserved properly. Freezing does not significantly affect final yield or purity. If autologous plasma is unavailable, AB plasma or commercial plasma substitutes can be used. For blood collection, tubes containing sodium or lithium heparin are recommended.
8. How can GVHD risk be minimized?
Literature indicates that γδT cells pose a lower risk of graft-versus-host disease (GVHD) due to their MHC-unrestricted recognition and immune-modulatory subtypes. Our expansion system primarily promotes Vδ2+ γδT cells, which can secrete IL-10 and TGF-β, helping suppress inflammatory responses while maintaining anti-tumor activity. Alternatively, αβT cells can be depleted using magnetic beads to further reduce GVHD risk.
9. If γδT cells reach 90% purity, what are the remaining cells? Are they αβT cells?
The remaining cells are typically CD3+ T cells. To further characterize the residual population, CD4 and CD8 surface markers can be assessed.
10. Which initial sample parameters affect the final purity of γδT cells?
Donor health, blood quality, anticoagulant type, seeding density, and culture conditions all influence the final outcome. While initial γδT percentages are typically below 5%, the YOCON γδT Expansion Kit is designed to consistently induce high-purity γδT cells regardless of the starting phenotype.
11. What is the typical killing efficiency of γδT cells in functional assays?
In a 20-hour co-culture assay using day-14 γδT cells and K562 tumor targets, killing efficiency reached 64% at a 20:1 E:T ratio and 93% at a 40:1 ratio.
