T-Cell Expansion Medium (Serum-Free)
MCE T-Cell Expansion Medium (Serum-Free) is a specialized culture medium formulated for the in vitro cultivation and robust expansion of human T lymphocytes without the need for serum.
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Storage :
T-Cell Basal Medium: 4°C, 1 year.
T-Cell Activation Reagent (200×): -20°C, 1 year. Avoid repeated freeze/thaw cycles.
- Manual (4895KB)
- COA
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Description & Advantages
MCE T-Cell Expansion Medium (Serum-Free) is a specialized culture medium formulated for the in vitro cultivation and robust expansion of human T lymphocytes without the need for serum.
This medium provides all essential nutrients required for T cell proliferation and supports the efficient expansion of T cells derived from peripheral blood mononuclear cells (PBMCs). It is also well-suited for the activation and expansion of T cells in culture. The expanded T cells are ideal for a wide range of in vitro applications, including co-culture experiments with tumor organoids for drug efficacy testing and high-throughput screening.
Protocol
Preparation of Complete T-Cell Expansion Medium
Prepare the complete T-Cell Expansion Medium by mixing the components below thoroughly. Keep the prepared medium on ice until use. Perform all steps under sterile conditions.
| Reagent | 10 mL | 20 mL | 50 mL |
|---|---|---|---|
| T-Cell Basal Medium | 9.85 mL | 19.7 mL | 49.25 mL |
| T-Cell Activation Reagent (200×) | 50 μL | 100 μL | 250 μL |
| FBS (MCE Cat. No.: HY-T1000) | 100 μL | 200 μL | 500 μL |
Note: For robust T-cell expansion, the Manual recommends adding FBS to the reconstituted T-cell expansion medium.
T-Cell Expansion Culture Setup Using PBMCs
1. PBMC Preparation: Isolate peripheral blood mononuclear cells (PBMCs) from fresh blood or thaw previously cryopreserved PBMCs.
2. Cell Counting: Resuspend PBMC-derived T cells in PBS and perform a viable cell count.
3. Centrifugation: Centrifuge the cells at 800 × g for 3 min and discard the supernatant.
4. Cell Resuspension and Seeding: Resuspend viable cells in complete T-Cell Expansion Medium and adjust the cell concentration to 1 × 106 cells/mL, or as required by the experimental design. Seed the cells into appropriate culture plates or petri dishes.
5. Incubation: Incubate at 37°C in a humidified incubator with 5% CO2.
6. Medium Replacement: Every 2-3 days, remove half of the spent medium and replace it with fresh complete T-Cell Expansion Medium.
7. Harvesting: T cells are typically ready for harvest after 10-14 days of culture.
8. Long-Term Expansion: For long-term expansion (>15 days), collect the T cells and resuspend them in fresh T-Cell Expansion Medium to continue culture.
Storage
T-Cell Basal Medium: 4°C, 1 year.
T-Cell Activation Reagent (200×): -20°C, 1 year. Avoid repeated freeze/thaw cycles.
Attention
1. This product is a sterile solution, pay attention to aseptic operation to avoid contamination.
2. This product is for R&D use only, not for drug, household, or other uses.
3. For your safety and health, please wear a lab coat and disposable gloves to operate.
Components
| Cat. No. | Components | HY-K3110-100 mL | HY-K3110-500 mL | Storage |
|---|---|---|---|---|
| HY-K3110-A | T-Cell Basal Medium | 100 mL | 500 mL | 4°C, 1 year. |
| HY-K3110-B | T-Cell Activation Reagent (200×) | 0.5 mL | 2.5 mL | -20°C, 1 year. Avoid repeated freeze/thaw cycles. |
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