Bambanker™ Direct: Freeze Hybridomas Straight from Culture, No Centrifugation
Bambanker™ Direct is a serum-free cell freezing medium that goes straight into your culture. Instead of pelleting cells and resuspending them in freezing medium, you add an equal volume of Bambanker™ Direct to the cells in their own medium, mix gently, aliquot and freeze at -80°C. It was developed for hybridomas, which are easily stressed by repeated handling and can lose viability or drift after freeze-thaw cycles, and for high-throughput work where spinning down dozens of clones is not practical.
Cutting out centrifugation, aspiration and resuspension reduces both hands-on time and mechanical stress on the cells. The medium is free of fetal bovine serum and antibiotics and protects cells through unregulated cooling, so there is no need for a programmable freezer. Vials can stay at -80°C or move to liquid nitrogen. Because it works in the culture vessel, Bambanker™ Direct suits freezing clones from multi-well plates during monoclonal antibody development as well as routine banking of suspension and adherent lines.
Key Benefits:
- Faster Workflow: No centrifugation. Add Bambanker™ Direct 1:1 to the culture and freeze. There is no pelleting, aspiration or resuspension step.
- Gentle on Cells: Less handling. Fewer manipulation steps mean less mechanical stress for fragile hybridomas and suspension cells.
- High-Throughput Ready: Built for many clones. Freeze clones directly from multi-well plates when screening or scaling up monoclonal antibody producers.
- Simple Storage: Direct to -80°C. No controlled-rate freezing. Store vials at -80°C or transfer to liquid nitrogen.
- Ready to Use: No preparation. Supplied as a complete solution, with nothing to dilute or mix before use.
Choosing the Right Bambanker™
Every Bambanker™ medium is ready to use, free of fetal bovine serum and antibiotics, and freezes cells directly at -80°C. They differ in what they are built to protect.
| Catalogue No. | Product | Best for | Formulation | Freezing workflow | Pack sizes |
|---|---|---|---|---|---|
| K1-0666 K1-0664 K1-0662 K1-0661 |
 Bambanker™ (Standard)View Product |
Routine and primary cell lines, tissue pieces, organoids and spheroids | Serum-free, 10% DMSO | Pellet, resuspend, freeze at -80°C | 10 mL, 20 mL, 5 × 20 mL, 100 mL |
| K1-0668 |  Bambanker™ DirectYou are here |
Hybridomas, fragile suspension cells, high-throughput clone banking | FBS-free (contains bovine serum-derived components) | Add 1:1 to cells in medium, freeze at -80°C. No centrifugation | 20 mL |
| K1-0676 |  Bambanker™ DMSO FreeView Product |
Cells that are damaged or differentiated by DMSO | Serum-free, DMSO-free | Pellet, resuspend, freeze at -80°C | 20 mL |
| K1-0672 K1-0673 |
 Bambanker™ hRMView Product |
Human and primate ES and iPS cells | Serum-free, xeno-free, human serum albumin, 10% DMSO | Pellet, resuspend, freeze at -80°C | 20 mL, 100 mL |
Technical Insights
One Step Shorter: The Bambanker™ Direct Workflow
Bambanker™ Direct is added straight to the cells in culture medium, removing the centrifugation and resuspension steps of the standard protocol.
| Step | Standard Bambanker™ | Bambanker™ Direct |
|---|---|---|
| 1 | Centrifuge cells and remove the medium | Add an equal volume of Bambanker™ Direct to the cells in medium |
| 2 | Resuspend the pellet in Bambanker™ | Pipette gently to mix |
| 3 | Transfer to cryovials | Transfer to cryovials |
| 4 | Freeze at -80°C | Freeze at -80°C |
Freezing suspension cells
- Check growth phase: Use cells in logarithmic growth phase for the best post-thaw viability.
- Add: Add a volume of Bambanker™ Direct equal to the volume of culture medium.
- Mix: Pipette gently until the suspension is uniform.
- Freeze: Transfer to cryovials and place directly at -80°C.
Freezing adherent cells
- Detach: Remove the medium, rinse twice with PBS and detach the cells with your usual dissociation reagent.
- Add: Add a volume of Bambanker™ Direct equal to the volume of detachment solution.
- Mix and freeze: Pipette gently to mix, transfer to cryovials and freeze at -80°C.
Thawing
- Thaw quickly: Warm the vial rapidly at 37°C.
- Dilute and wash: Suspend the cells in culture medium, centrifuge and remove the supernatant.
- Culture: Resuspend in fresh medium and seed.
Cell Lines Frozen Successfully with Bambanker™ Direct
Although it was designed with hybridomas in mind, Bambanker™ Direct has been used successfully with a broad range of suspension and adherent cell lines.
| Cell line | Origin |
|---|---|
| C2C12 | Mouse skeletal muscle myoblast |
| Daudi | Human B lymphoblast (Burkitt’s lymphoma) |
| HEK293 / HEK293T | Human embryonic kidney |
| HeLa / HeLa S3 | Human cervical cancer |
| HepG2 | Human liver cancer |
| Jurkat | Human T-cell leukaemia |
| K562 | Human chronic myeloid leukaemia |
| KATO | Human gastric cancer |
| MDCK | Dog kidney epithelial |
| NIH3T3 | Mouse embryonic fibroblast |
| OKT4 | Mouse hybridoma |
| OP9 | Mouse bone marrow stromal |
| P3U1 | Mouse myeloma |
| PC12 | Rat adrenal phaeochromocytoma |
| SNL | Mouse embryonic fibroblast feeder |
| Mouse ES cells | Mouse embryonic stem cells |
| MEF | Mouse embryonic fibroblast |
| Vero | African green monkey kidney |
Bambanker™ Direct belongs to the Bambanker™ family, which has been cited across more than 100 cell and sample types and adopted by the JCRB Cell Bank. See the standard Bambanker™ page for the full validation data and publication list.
Bambanker™ Direct Frequently Asked Questions
1. How is Bambanker™ Direct different from standard Bambanker™?
Standard Bambanker™ replaces the culture medium: you pellet the cells and resuspend them in it. Bambanker™ Direct is added to the medium at a 1:1 ratio, so the cells never need to be spun down before freezing.
2. Is it only for hybridomas?
No. It was developed for hybridomas and high-throughput clone banking, but it has been used with many suspension and adherent lines, including Jurkat, K562, HEK293T, HeLa and NIH3T3. See the table above.
3. Can I use it for adherent cells?
Yes. Detach the cells as usual, then add Bambanker™ Direct in an equal volume to the detachment solution, mix and freeze.
4. Do I need a programmable freezer?
No. Vials go straight into a -80°C freezer and can later be moved to liquid nitrogen if needed.
5. Does it contain serum or animal-derived components?
It is free of fetal bovine serum and antibiotics. The manufacturer’s manual notes that it contains bovine serum-derived compounds, so it is not suitable where an animal-component-free or xeno-free medium is required. For human ES and iPS cells in xeno-free workflows, use Bambanker™ hRM.
6. How should I store the medium?
Store at 2–10°C. Shelf life is 2 years from the date of manufacture.
7. What growth phase should the cells be in?
Freeze cells in logarithmic growth phase. Cells frozen from older or overgrown cultures recover less well.
8. Is it made to GMP?
The Bambanker™ series is manufactured in GMP-compliant facilities. For research use only. Not for use in humans or for diagnostic or therapeutic procedures.
Specifications
- Product type: Serum-free cell freezing medium for direct addition to culture
- Formulation: Ready to use. Free of fetal bovine serum and antibiotics. Contains bovine serum-derived components
- Use: Add 1:1 to cells in culture medium or detachment solution
- Suitable for: Hybridomas, suspension and adherent cell lines, high-throughput clone banking
- Freezing: Directly at -80°C, with no centrifugation or programmed freezing
- Storage of frozen cells: -80°C or liquid nitrogen (-196°C)
- Storage of medium: 2–10°C
- Shelf life: 2 years from date of manufacture
- Pack size: 20 mL (K1-0668)
- Quality: Manufactured in GMP-compliant facilities
- Intended use: For research use only. Not for use in humans or for diagnostic or therapeutic procedures.