Cryopreservation of mammalian cells is incredibly useful and frequent in biological research. The fear of losing a cell line due to contamination or incubator failure is typically the incentive for prioritizing cell archival preservation after obtaining or creating a new cell line.
Once moved from growing media to freezing medium, the cells are usually frozen at a regulated pace and preserved in liquid nitrogen vapor or at -130 °C in a mechanical deep freeze. Although freezing a cell line is a standard technique, issues emerge when proper freezers are unavailable or when undesirable variables are added due to serum, extra-wash, or intricate freezing algorithms.
- Enables long-term cell storage at -80 °C or -196 °C
- Improved cell survival and viability after freezing
- Ready-to-use freezing media–no preset or sequential freezing required
- Serum-free, with no chance of contamination
- Produced in GMP-compliant facilities (further info)
- Suitable for all known cell lines.
- Long shelf life
Image Credit: NIPPON Genetics EUROPE GmbH
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Reasons to trust Bambanker™ cell freezing medium
- Secure serum-free medium | No possibility of contamination from FBS, and consistent product quality
- Time-saving and easy freezing at -80 °C possible - No step-by-step freezing required
- Highest cell viability and recovery > 90%: Keep the cells safe
- Easy and quick protocol: Simply add and freeze
- GMP quality: Manufactured in a GMP-compliant facility
Save time while saving the cells
Cryopreservation of cells at -80 °C in liquid is made possible by the cell freezing media Bambanker™, which eliminates the need for a separate, costly ultralow freezer as well as time-consuming, complex controlled freezing procedures. Cells are simply harvested, aspirated, resuspended in Bambanker™, transferred to a cryovial, and stored at -80 °C. There is no need for sequential or planned freezing.
Bambanker™ is a ready-to-use, serum-free medium that can be stored in the refrigerator for up to two years. Since Bambanker™ freezing medium comes in handy 20 ml vials, it is perfect for individual lab members to use.
Bambanker™, a cutting-edge cell freezing medium, is now the market leader in Japan and is distinguished by several published articles using extremely sensitive cell lines worldwide.
Image Credit: NIPPON Genetics EUROPE GmbH
Serum adds variation to long-term storage
Bambanker™ goods are created without serum. Cryopreservation media containing sera have the drawback of variable recovery rates and an unknown composition. The reproducibility of investigations with cells frozen in a serum-containing medium can be influenced by lot-to-lot variation in the serum, as the content and quantity of proteins and other biological molecules vary with each batch of serum.
This may cause complications when thawing and utilizing cells from serum-containing media. Bambanker™’s carefully established ingredients ensure that cells stored at different periods behave and recover identically.
Universal – Even for preservation of stem cells
All common cell lines can result in a large number of viable cells following thawing. Recovery rates for even sensitive cells are significantly greater than in regular media. The JCRB cell bank successfully stores over 1,400 distinct cell lines using Bambanker™.
High quality – Manufactured in GMP-compliant facilities
All items of the series are made in facilities compatible with current good manufacturing practices (GMP).
Flexible
There are four different versions of Bambanker™. All Bambanker™ types do not contain fetal bovine serum (FBS) and have a defined, consistent composition. They also do not contain antibiotic or antifungal ingredients.
The four different types of Bambanker™ are recommended depending on specific needs.
Applications
Bambanker™ prevents undesired differentiation
Cell viability and ALP staining of pluripotent stem cells. Upper row: A great number of cells are detected two days after thawing. The cells show no morphological change after thawing. Lower row: Bambanker™ does not cause cell differentiation as all stem cells frozen down are still producing high levels of alkaline phosphatase, a reporter for pluripotent stem cells. Image Credit: NIPPON Genetics EUROPE GmbH
Comparison of Bambanker™ with another cryopreservation medium for the cultivation of 1,400 different cell lines
The JCRB cell bank has roughly 1,400 distinct cell lines. Poor survival rates after thawing frozen cell lines (KHYG-1, KAI3, HL60, OVMANA) led researchers to test Bambanker™ and compare it to the previously used preservation medium. Cell growth efficiency after thawing was tested between the commercially available preservation medium and Bambanker™.
All cultured cells were extracted from the logarithmic growth phase. A storage tube was filled with roughly 1×106 cells and 1 ml of preservation medium. Cells were kept at -80 °C for two weeks. The frozen cells were thawed in a 37 °C water bath and then incubated in a 96-well plate with 5% CO2. Viable cell counts were determined daily.
Image Credit: NIPPON Genetics EUROPE GmbH
The survival rate of four cell lines (KHYG-1, HL60, KAI3, and OVMANA) after thawing is quite low when using commercially available products and Bambanker™. After thawing, Bambanker™ enhanced cell proliferation in all four cell lines, outperforming the existing commercial product. Since 2014, the JCRB cell bank has used Bambanker™ for its cell lines.
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Bambanker Direct
- Enables long-term cell storage -80 °C or -196 °C
- Ideal for hybridoma cells and HTP-applications
- Bambanker Direct can be administered straight to the cells - no washing or centrifugation required
- Improved cell survival and viability after thawing
- Ready-to-use freezing media–no programmed or sequential freezing required
- Serum-free, with no chance of contamination
- Produced in GMP-compliant facilities (further info)
- Long shelf life
Image Credit: NIPPON Genetics EUROPE GmbH
Details
Bambanker™ Direct for hybridomas
Certain cell types, such as hybridomas, are sensitive to “over” manipulations. These cells frequently die or differentiate abnormally following long-term storage and freeze-thaw cycling. Bambanker™ Direct was designed for cells that require precise environmental control. By skipping the centrifugation stage, cells can now be frozen for long-term preservation directly after adding the new cryoprotectant.
Simply apply Bambanker™ Direct one-to-one to the cells in media and freeze. The Direct version, like the original Bambanker™, is serum-free, making it ideal for cells that can be sensitive to animal-derived serums.
Procedure for the cryopreservation of cells
Image Credit: NIPPON Genetics EUROPE GmbH
High quality – Manufactured in GMP-compliant facilities
Bambanker™ products are made in facilities that follow current good manufacturing practices (GMP). The products are subject to a rigorous quality management system that provides consistency, dependability, and excellent quality on which customers can rely.
Cell lines
Cell lines successfully stabilized by Bambanker™ Direct:
- NIH3T3 (mouse embryonic skin cell line)
- OKT4 (mouse hybridoma)
- OP9 (mouse myeloid stromal cells)
- PC12 (rat adrenal medulla cell line)
- SNL (mouse embryonic fibroblasts)
- ES (mouse embryonic stem cells)
- MEF (mouse embryonic fibroblasts)
- Vero (African green monkey kidney cell line)
- C2C12 (mouse skeletal muscle cell line)
- Daudi (human B cell line)
- HEK293; HEK293T (human embryonic kidney cell line)
- HeLa/HeLa S3 (human cervical cancer cell line)
- HepG2 (human liver cancer cell line)
- Jurkat (human leukemia T cell line)
- K562 (human chronic myelogenous leukemia cell line)
- KATO (human gastric epithelial cell line)
- MDCK (canine kidney tubular epithelial cell line)
- OP9 (mouse myeloma stroma cells)
- P3U1 (mouse myeloma cell line)
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Bambanker hRM
Serum-free cryopreservation media - For ES and iPS cells
- Ready-to-use freezing media–no programmed or sequential freezing required
- Serum-free, with no chance of contamination
- Produced in GMP-compliant facilities (further info)
- Long shelf life
- Ideal for the cryopreservation of ES and iPS cells
- Made using human serum albumin–no other animal components
- Enables long-term cell storage at -80 °C or -196 °C
- Improved cell survival and viability after thawing
Image Credit: NIPPON Genetics EUROPE GmbH
Details
A new hope for the cryopreservation of ES and iPS cells
Primate embryonic stem cells are particularly sensitive to cryopreservation, posing several challenges compared with murine or other cells. Currently, the vitrification procedure is deemed appropriate for primate ES/iPS cells; the slow-freezing method with DMSO has proved popular for a wide range of cell lines.
Vitrification is the fast cooling of frozen medium to a glass-like, crystalline state. The vitrification procedure demands precise timing and a high level of competence, yet it can still provide unsatisfactory results. Furthermore, this approach demonstrates sensitivity to dry ice transportation. To overcome these issues, the novel freezing medium, Bambanker™ HRM, was created.
Image Credit: NIPPON Genetics EUROPE GmbH
What makes Bambanker™ HRM so unique?
The Bambanker™ HRM cell media does not include bovine serum albumin, which could lead to cell differentiation. Furthermore, there is no animal-derived material to increase the storage and survival of primate ES/iPS cells while simplifying the technique.
Cell freezing without serum
No serum is used in any Bambanker™ products. Cryopreservation media that contain sera have the drawback of inconsistent recovery rates and unclear composition. Since the content and quantity of proteins and other biological molecules vary with each batch of serum, lot-to-lot variation in the serum could impact the reproducibility of research with frozen cells in a serum-containing medium.
Freezing and using cells from such serum-containing media can cause problems. Users can be sure that cells stored at different periods will all function and recover similarly since every element in Bambanker™ is carefully specified.
Applications
Customer product feedback
Comparison of the cryopreservation efficiency for human iPS cells
Human iPSC 201B7 cells were cryopreserved using Bambanker™ HRM, a standard vitrification medium, or a 10% DMSO/culture mixture. After three days, the cells were thawed and plated according to the procedure.
In the case of a 10% DMSO medium, only a few colonies could be retrieved, indicating a low survival rate, but with Bambanker™ HRM, massive colonies of about the same size as those obtained with vitrification freezing preservation solution were recovered. These results show that Bambanker™ HRM can achieve the same storage efficiency as the vitrification preservation technology, with the added benefit of easy handling.
Data kindly provided by US corporation Hasumi International Research Foundation, Tokyo Research Center, Japan. Image Credit: NIPPON Genetics EUROPE GmbH
Comparison of cryopreservation efficiency for the common marmoset fibroblast cells intended for the iPS cell induction
For fibroblasts from common Marmoset, the storage effectiveness of Bambanker™ and two other commercial preservative solutions (supplier T, supplier S) was tested. In a 6 cm Petri dish, fibroblasts were cultivated until they reached a confluency of 90–100%. The cells were frozen at -80 °C for two months.
Photomicrographs were obtained for each preservation solution 24 and 48 hours after thawing. The proportion of dead cells was calculated. Bambanker™ HRM had the best survival rate, followed by Supplier Ts storage solution.
The storage solution from supplier Ss has the lowest survival rate. The only cells that demonstrated a significant quantity to be utilized directly for iPS cell induction were those that were kept with Bambanker™ HRM.
Data kindly provided by Primate Research Institute, University of Kyoto, Molecular Physiological Research Department. Image Credit: NIPPON Genetics EUROPE GmbH
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Bambanker DMSO Free
Serum-free cryopreservation media - For most sensitive cells
- Serum-free, with no chance of contamination
- Produced in GMP-compliant facilities (further info)
- Suitable for all known cell lines
- Long shelf life
- DMSO-free--for the most sensitive cells
- Enables long-term cell storage at -80 °C or -196 °C.
- Improved cell survival and viability after thawing
- Ready-to-use freezing media–no preset or sequential freezing required
Image Credit: NIPPON Genetics EUROPE GmbH
Details
No more DMSO – For the most sensitive cells
Most freezing reagents contain DMSO to help prevent the production of ice crystals, which can be harmful to cells. However, DMSO is cytotoxic and can impair the survival rate of some sensitive cell types.
Bambanker DMSO Free is formulated without DMSO. Instead, it employs a special composition to prevent the production of ice crystals. This makes Bambanker DMSO Free ideal for cell lines that have shown sensitivity to DMSO under long-term preservation settings.
High quality – Manufactured in GMP-compliant facilities
All Bambanker products are made in facilities that follow current good manufacturing practices (GMP). The products are subjected to a rigorous quality management system that assures consistency, dependability, and superior quality.
Cell lines
Cell lines
The following cell lines were effectively frozen using Bambanker DMSO-free:
- Jurkat (Floating cells)
- PC12 (Adrenal Pheochromocytoma, Rat) (Adhesive cells)
- HepG2 (Adhesive cells)
- Hela S3 (Epithelioid Carcinoma, Cervix, Human) (Adhesive cells)
- HEK293T (Transformed Primary Embryonal Kidney, Human) (Adhesive cells)
- OKT4 (Floating cells)
- Vero P4 (Kidney, African Green Monkey (Adhesive cells)
- MDCK (Kidney, Canine)(Adhesive cells)
- NIH3T3 (Embryonic Fibroblast, Mouse) (Adhesive cells)