login_enqueue_scriptshooks. This notice was triggered by the
font-awesomehandle. Please see Debugging in WordPress for more information. (This message was added in version 3.3.0.) in /var/www/vhosts/nippongenetics.eu/stage.nippongenetics.eu/wp-includes/functions.php on line 5311
Serum-free freezing media for long storage of 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. 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.
The cell freezing media BambankerTM permits cryopreservation of cells at -80°C (or in liquid nitrogen), obviating the need for an additional and expensive ultralow freezer and avoiding time consuming and complicated controlled freezing protocols. Simply (1) harvest cells, (2) aspirate medium, (3) resuspend in BambankerTM (4) transfer to a cryovial and (5) store at -80°C. No programmed or sequential freezing is required! BambankerTM 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 BambankerTM freezing medium ideal for use by individual members of your lab. Today this innovative cell freezing medium BambankerTM is the market leader in Japan and characterized by many different published articles with very sensitive cell lines all over the world.
All BambankerTM 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 BambankerTM is precisely defined so that you can be confident that cells stored at different times will all behave and recover similarly.
All common cell lines can be preserved resulting in a high number of intact cells after thawing. Recovery rates of even sensitive cells is much higher compared to regular media. For example the JCRB cell bank stores over 1,400 different cell lines with great success with BambankerTM.
There a four different version of BambankerTM:
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: BambankerTM 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.
The JCRB cell bank handles approximately 1,400 different cell lines. A low survival rate after thawing frozen cell lines (KHYG-1, KAI3, HL60, OVMANA) has let them to test BambankerTM and compare it to the up to then used preservation medium for the four cell lines. The growth efficiency after thawing was compared for cells stored with the currently used commercially available preservation medium and BambankerTM.
All cultured cells were harvested in the logarithmic growth phase. 1 ml preservation media was added to approximately 1 x 106 cells in a storage tube. The cells were stored for 2 weeks at -80°C. The frozen cells were thawed in an 37°C water bath and incubated at 37°C and 5% CO2 in a 96-well plate. Every day the viable cell number was determined.
The survival rate after thawing of the four cell lines (KHYG-1, HL60, KAI3, OVMANA) is with the currently used commercially available product and Bambanker™ very low. However, after thawing, all four cell lines cell proliferation was improved with Bambanker™ when compared to the currently used commercial product.
→ The JCRB cell bank has been using BambankerTM since 2014 for all their cell lines.
→ Download here AppNote for more Information
Successful frozen cells
The JCRB cell bank stores over 1,400 different cell lines with great success with BambankerTM.
Yes, the only Bambanker that does not contain DMSO is Bambanker DMSOfree (BBF01, BBF02)
Yes, you do not need a washing step.
As long as single cells are used, there is no problem. We do not have experience for whole organism.
This is no problem.
1 ml is recommend for 10^6 cells, you can make different downsizing 0.5 for 5×10^5 and so on.
Yes. But the low amount of 10^6 cells can make a problem in the RNA Isolation and will lead to a low amount of RNA.
You can use normal Bambanker for animal tissue. Also we would recommend to amke the tissue finer than 20 mg. Also you can place the tissue in Bambanker at 4°C for 10 minutes before freezing it.
For further information, please take a look in these papers:
This is no problem, as long as the cells keep healthy.
This is no problem. Use it directly.