Bambanker hRM for ES and iPS cells
Human/primate embryonic stem cells and induced-pluripotent stem cells are extremely sensitive to cryopreservation, which presents many difficulties when compared cells from mice and other animal sources. 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 requires impeccable timing, a high level of skill, and still can yield poor results when everything is done right. In addition, this method shows sensitivity to dry ice transportation. To address these problems, a new freezing method for human cells has been developed and based upon the primate-specific Bambanker hRM cryopreservation reagent. What makes it so special is the omission of bovine serum albumin (which can sometimes lead to cell differentiation) and any animal-derived material — it’s “xeno-free” — which can improve the storage and survival rates of a broad range of primate/human cells lines, primary cells and stem cells — while greatly simplifying the freezing protocol. Here are some Technical Notes demonstrating the advantages:
Bambanker is easy to use – and better for your cells!
Cryopreservation of mammalian cells is extremely valuable and very 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 freezer space is not available, or when unwanted variables are introduced by the presence of serum, extra washes, or complicated freezing algorithms.
These problems have been resolved by the family of Bambanker Cell Freezing Media reagents. 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:
- harvest and prepare cells
- resuspend in one of four types of Bambanker
- transfer to a cryovial
- immediately freeze at -80°C
No programmed or sequential freezing is required! All Bambanker reagents are free from serum — delivered ready-to-use — and can be kept in the refrigerator for up to two years.
Higher number of intact, healthy, and undifferentiated cells after thawing
How much are your cells worth? The family of Bambanker freezing reagents offer the unique combination of simplicity and high survivability. The results can be miraculous. These products have been used extensively across the globe. Developed in Japan, Bambanker reagents have been validated on well over 1,000 different cell lines at the JCRB. You can also see these recent published references as a testament to its successful use in labs. Recovery rates, even of sensitive cells, are much higher compared to regular cell freezing media and techniques. The vast majority of cell types show survival greater than 50%, with many at 90% or more. Even hematopoietic stem cells pose no problem.
Unwanted cell differentiation can also be an issue with long-term storage of frozen cells. All Bambanker reagents are designed to minimize this issue. And for particularly troublesome cells, Bambanker DMSO-Free is specially formulated to prevent such issues that can be associated with DMSO-containing cryopreservation reagents. 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 animal-derived whole serum have the disadvantage of fluctuations in recovery rates and undefined composition. Reproducibility of experiments with cells frozen in a serum-containing cryopreservation medium may 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. Problems may arise when thawing and using cells stored with such serum-containing freezing reagents. But you’ll be able to breathe easy with Bambanker reagents: every ingredient is precisely defined, including the use of recombinant serum albumin proteins, so that you can be confident that Bambanker-preserved cells 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 10^8 cells), hematopoietic stem cells. The left picture shows the cells before freezing and the right picture shows the cells after freezing and thawing.
Bambanker HRM Cell Freezing Medium
|Part Numbers||BBH01, BBH03|
|Serum Albumin||Yes, recombinant human serum albumin|
|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|