This PBMC Cryopreservation Medium is a ready‑to‑use serum‑free cryoprotective solution specially designed for cryopreservation of Peripheral Blood Mononuclear Cells (PBMC). Formulated with an optimized ratio of permeable and non‑permeable cryoprotectants, it balances ice crystal damage and osmotic injury during cooling. Key features are as follows:
1. Ready‑to‑use, no on‑site preparation required;
2. Minimal cell damage during freezing and thawing; post‑thaw cell viability ≥90% (subject to actual test results);
3. Post‑thaw cell phenotypes (CD3/CD4/CD8/CD19, etc.) remain consistent with pre‑freezing status, with no significant impairment to proliferation and functional activity;
4. Suitable for short‑term storage at −80 °C and long‑term storage in liquid nitrogen (−196 °C) of PBMC and various suspended immune cells.
Main Components:
| Component Category | Description |
| Cryoprotectant | DMSO (final concentration ≤10%) |
| Serum replacement components | Animal-origin-free, protein hydrolysate / albumin substitutes |
| Buffer system | Buffered salt system to maintain physiological pH |
| Nutritional and stabilizing components | Amino acids, carbohydrates, antioxidants, etc. |
| Antibiotics | None (to be added by user if required) |
Instructions for Use:
1. Cell Cryopreservation
1.1 Harvest freshly isolated or cultured PBMC, count cells and confirm viability ≥90% (Trypan Blue or AO/PI method).
1.2 Centrifuge at 300 × g for 5–10 min at room temperature and discard the supernatant.
1.3 Resuspend the cell pellet with pre‑chilled (2–8 °C) PBMC Cryopreservation Medium, adjust cell density to 1×10⁷–1×10⁸ cells/mL, and mix gently.
1.4 Aliquot 1 mL into each sterile cryovial, cap tightly and label.
1.5 Controlled cooling: Place cryovials in a controlled‑rate cooling container (with isopropanol) at −80 °C for ≥4 h, or follow gradient cooling: 4 °C for 10 min → −20 °C for 30 min → overnight at −80 °C.
1.6 Long‑term storage: Transfer cryovials to liquid nitrogen (−196 °C) and record storage locations.
2. Cell Thawing
2.1 Remove the cryovial from liquid nitrogen and thaw rapidly in a 37 °C water bath with gentle agitation until completely melted within 1–2 min.
2.2 Disinfect the vial surface with 75% ethanol. Slowly transfer the cell suspension dropwise into a centrifuge tube containing pre‑warmed complete medium while swirling gently.
2.3 Centrifuge at 300 × g for 5 min and discard the supernatant to remove cryoprotectant.
2.4 Resuspend cells with fresh medium for culture or downstream assays as required.
2.5 It is recommended to count and test cell viability 24 h after thawing to verify recovery performance.
Tip: This protocol is for reference only. Users may optimize it according to their own workflows (e.g., CAR‑T / NK processes, whole‑blood isolation protocols).
Precautions:
1. For Research Use Only. Not for clinical diagnosis, therapy or human infusion.
2. All operations must be performed aseptically in a biosafety cabinet to prevent microbial contamination.
3. This formulation contains DMSO, which is volatile and irritating to skin. Operate under ventilation and avoid direct contact with skin and mucous membranes.
4. This product is ready‑to‑use. Do not dilute, add extra DMSO or subject it to repeated freeze‑thaw cycles.
5. It is recommended to use up the product once opened. If aliquoting is needed, perform aseptically and store the aliquots short‑term at 2–8 °C.
6. Do not use if the liquid appears turbid, precipitated, discolored or otherwise abnormal; contact your supplier.
7. Dispose of waste liquid and contaminated consumables in accordance with biohazard waste regulations.
FAQs:
1. What to do if post‑thaw viability is low?
Suggestion: Verify the cooling procedure (avoid direct rapid freezing at −80 °C); ensure thawing is completed within 1–2 min in a 37 °C water bath; check cell quality before freezing.
2. What to do if cells aggregate after thawing?
Suggestion: Add cell suspension slowly and mix gently during dilution; filtration through a 40 μm cell strainer may be applied if needed.
3. Can it be used for T cells / NK cells?
Suggestion: Optimized for PBMC and compatible with T, NK and other immune cells. Dedicated freezing medium is recommended for large‑scale T/NK manufacturing.
4. Can it be used for therapeutic / clinical purposes?
Suggestion: This research‑grade product is not for clinical use. Therapeutic‑grade products are required for clinical applications.
