Save time while saving your cells
Cryopreservation of mammalian cells is extremely valuable and common in biological research. Fear of losing a cell line to contamination or incubator failure is frequently the impetus for making archival storage of cells a high priority after receiving or generating a new cell line. While there are numerous protocols for freezing cells, all involve the use of a ‘freezing medium’ containing a cryoprotectant. Once transferred from growth media to freezing medium, the cells are usually frozen at a controlled rate and stored in liquid nitrogen vapor or at -130°C in a mechanical deep freeze. Although freezing a cell line is a commonly performed procedure, problems arise when suitable freezers are not available, or unwanted variables are introduced by the presence of serum, extra-wash or complicated freezing algorithms.
These problems have been resolved by Bambanker Cell Freezing Media. The unique technology of Bambanker permits cryopreservation of cells at -80°C (or in liquid nitrogen), obviating the need for an additional and expensive ultra-low freezer and avoiding time consuming and complicated controlled freezing protocols. Simply i) harvest cells, ii) aspirate medium, iii) resuspend in Bambanker iv) transfer to a cryovial and v) store at -80°C. No programmed or sequential freezing is required! Bambanker is a serum-free cryopreservation medium that is delivered ready-to-use and can be kept in the refrigerator for up to two years. Convenient 20 ml bottles are available, making Bambanker Freezing Medium ideal for use by individual members of your lab.
- Up to 3x Better Recovery Rates
- No Step-wise Freezing
- -80°C or Liquid Nitrogen Storage
- 1-Step Protocol for Fragile Cells
Bambanker HRM for ES and iPS cells
Primate embryonic stem cells and induced-pluripotent stem cells are extremely sensitive to cryopreservation which presents many difficulties when compared to murine or other cells. At present, the vitrification method is considered adequate for primate ES/iPS cells although the slow-freezing method using DMSO has been popular for a wide variety of cell lines. Vitrification is the rapid cooling of the freezing media to a glass-like crystalline state. The vitrification method requires impeccable timing, a high level of skill, and still can yield poor results. In addition, this method shows sensitivity to dry ice transportation. To address these problems, a new freezing medium called Bambanker HRM has been developed. What makes it so special is the removal of bovine serum albumin (which can sometimes lead to cell differentiation) and any animal-derived material “xeno-free”, both improve the storage and survivability of primate ES/iPS cells while greatly simplifying the protocol.
To prove the efficacy, primate ES cells were cryopreserved in either Bambanker HRM or a 10% DMSO/culture medium by slow-freezing or in a conventional vitrification mediums by quick-freeezing, and then subsequent storage in liquid nitrogen. After 3 days, the cells were thawed, according to protocol, and then plated. The different cryopreservation media were analyzed by the number of alkaline phosphatase-positive colonies as recovery points. The recovery points of Bambanker HRM directly from liquid nitrogen storage were twice as high compared with vitrification medium and four times that of 10% DMSO/culture medium. To simulate dry ice transportation, the cryopreserved primate ES cells above were put on dry ice for 24 hours after three days in liquid nitrogen, and then thawed and plated. The recovery points of Bambanker HRM still remained high, but those of vitrification were considerably lower.
Bambanker DMSO-FREE for sensitive cells
DMSO is added to most freezing reagents to help avoid formation of ice crystals, which of course harm the cells. However, DMSO is cytotoxic and can reduce the survival rate of certain sensitive cell lines. Bambanker DMSO Free is made without DMSO. Instead, it uses a unique formulation to avoid the formation of ice crystals. This makes Bambanker DMSO Free especially suitable for cell lines that prove to be sensitive to DMSO under long-term storage conditions.
Bambanker DIRECT for hybridomas
There are certain cell types, such as hybridomas, which exhibit increased sensitivity to “over” manipulations. These cells often exhibit higher death rates or unwanted differentiation after long-term term storage and freeze thaw cycling. It is for these types of cells that Bambanker DIRECT was created – to provide tight environmental control. By eliminating the centrifugation step, cells can now be frozen for long-term storage immediately upon addition of the novel cryoprotectant. Just add Bambanker DIRECT, one-to-one, with your cells in media and directly place in freezer. And like with the original Bambanker, the DIRECT version has no serum, so it is great for cells which animal-derived serum may be an issue.
Higher number of intact cells after thawing
How much are your cells worth? Bambamker Freezing Media offer the unique combination of simplicity and high survivability. The results are simply miraculous. This product has been used extensively in Asia, where it was first developed, with a broad range of cell types. See these recent published references as a testament to its use. Recovery rates, even of sensitive cells, are much higher compared to regular cell freezing media. The vast majority of cell types show survivability greater than 50% with many approaching 90% or more. Important cell lines like hybridomas can have as high as 100% recovery after long-term storage. Even hematopoietic stem cells pose no problem. So come join the growing number of labs protecting their cell lines with Bambanker.
Serum adds variation to long-term storage
All Bambanker products are produced with no serum. Cryopreservation media which contain sera have the disadvantage of fluctuations in recovery rates and undefined composition. Reproducibility of experiments with cells which were frozen in a serum-containing-medium, could be affected by lot-to-lot variation of the serum since the composition and concentration of proteins and other biological molecules varies with each batch of serum. This may result in issues when thawing and using cells from such serum-containing media. Breathe easy, as every ingredient of a Bambanker product is precisely defined so that you can be confident that cells stored at different times will all behave and recover similarly.
Cells were incubated as described in the protocols. Cell viability was determined after twelve months
Cell freezing experiment with HSC (8.8 x 108 cells), hematopoietic stem cells. The left picture shows the cells before freezing and the right picture shows the cells after freezing and thawing.
General Cells - Bambanker Cell Freezing Medium
Hybridoma Cells Bambanker DIRECT Cell Freezing Medium
ES and IPS Cells Bambanker HRM Cell Freezing Medium
DMSO-Free Bambanker DMSO Free Cell Freezing Medium
|Cyroprotectant||DMSO-based (except DMSO-free Bambanker)|
|Part Numbers||BB01, BB02, BB04, BBD01, BBH01, BBF01, BBF02|
|Cell Type Compatibility||All, adherent & suspension|
|Cell Storage Temp||-80°C or liquid nitrogen|
|Survival after 12 months||50% to 100% of cells|
|Storage||2 years at 4°C|