Cryopreservation media

Bambanker (20 ml)

Article number: BB03

20 ml serum-free freezing media for long storage of cells

 

Key Benefits

  • Enables long term cell storage at -80°C or -196°C
  • Improved cell recovery and viability after thawing
  • Ready-to-use freezing media – no programmed or sequential freezing required
  • Serum-free – no risk of contamination
  • Usable for all know cell lines
  • Long shelf life

Long-term storage of cultured 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.

 

Save time while saving your cells

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.

Freezing protocol of Bambanker

Serum adds variation to long-term storage

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.

 

Universal – even for preservation of stem cells

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.

 

Flexible

There a four different version of BambankerTM:

1. BambankerTM (Universal)

2. BambankerTM HRM

3. BambankerTM Direct

4. BambankerTM DMSO-Free

BambankerTM prevents undesired differentiation

 

No undesired differentiation with Bambanker

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.

 

Comparison of BambankerTM with another cryopreservation medium for the cultivation of 1,400 different cell lines

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.

 

Comparison of Bambanker with another cryopreservation medium

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.

Successfully frozen cell lines with Bambanker

  1. Hatoya, S. et al. Effect of co-culturing with embryonic fibroblasts on IVM, IVF and IVC of canine oocytes. Theriogenology 66, 1083–1090 (2006).
  2. Hikichi, T. et al. Differentiation potential of parthenogenetic embryonic stem cells is improved by nuclear transfer. Stem Cells 25, 46–53 (2007).
  3. Huang, Y. H., Yang, J. C., Wang, C. W. & Lee, S. Y. Dental Stem Cells and Tooth Banking for Regenerative Medicine. J. Exp. Clin. Med. 2, 111–117 (2010).
  4. Lechner, S. M. Interaktionen von Inseminatbestandteilen mit Epithelzellen und Leukozyten im Uterus des Rindes. (2008).
  5. Liu, D. et al. Relation between human decay-accelerating factor (hDAF) expression in pig cells and inhibition of human serum anti-pig cytotoxicity: Value of highly expressed hDAF for xenotransplantation. Xenotransplantation 14, 67–73 (2007).
  6. Mieno, S. et al. Effects of diabetes mellitus on VEGF-induced proliferation response in bone marrow derived endothelial progenitor cells. J. Card. Surg. 25, 618–625 (2010).
  7. Mieno, S. et al. Characteristics and Function of Cryopreserved Bone Marrow-Derived Endothelial Progenitor Cells. Ann. Thorac. Surg. 85, 1361–1366 (2008).
  8. Naito, H. et al. The advantages of three-dimensional culture in a collagen hydrogel for stem cell differentiation. J. Biomed. Mater. Res. – Part A 101, 2838–2845 (2013).
  9. Shimizu, Y. et al. Impaired tax-specific t-cell responses with insufficient control of HTLV-1 in a subgroup of individuals at asymptomatic and smoldering stages. Cancer Sci. 100, 481–489 (2009).
  10. Takata, Y. et al. Generation of iPS Cells Using a BacMam Multigene Expression System. Cell Struct. Funct. 36, 209–222 (2011).
  11. Tamai, Y. et al. Potential contribution of a novel Tax epitope-specific CD4+ T cells to graft-versus-Tax effect in adult T cell leukemia patients after allogeneic hematopoietic stem cell transplantation. J. Immunol. 190, 4382–92 (2013).
  12. Zaidi, S. K. et al. Runx2 deficiency and defective subnuclear targeting bypass senescence to promote immortalization and tumorigenic potential. Proc. Natl. Acad. Sci. U. S. A. 104, 19861–19866 (2007).

on 07/23/2018

Mit der Verwendung des Bambaker Einfriermediums konnten wir den prozentualen Anteil überlebender iPS-Zellen nach dem Auftauen wesentlich erhöhen. Wir sind mit diesem Produkt überaus zufrieden und verwenden es mittlerweile für alle Stammzellkulturen.

Translation by Nippon Genetics:

By using the Bambaker freezing medium, we were able to increase significantly the percentage of surviving iPS cells after thawing. We are very satisfied with this product and now we use it for all our stem cell cultures.

on 02/02/2018

Since many years we are working with primary cell cultures derived from high-grade brain tumors which are highly susceptible to environmental changes and differing culture conditions. Thus, freezing and thawing is a challenging process for these cells. According to our experiences, we could not avoid to lose 30% to 40% of living cells after each freezing/thawing cycle. In addition, the cells need a recovery time of at least five days after freezing in our previous freezing medium, obtained from other manufacturers. We’ve recently tested the Bambanker medium with striking results: recovery times of only two days of 80% of living and “healthy” cells. We observed this improvement of recovery time and proportion of living cells with all four cell lines, that we’ve tested so far. We are very satisfied with Bambanker and will continue to use it.

Translated by Nippon Genetics:

Wir arbeiten seit vielen Jahren mit primären Zellkulturen, die aus weitfortgeschrittenen Gehirntumoren gewonnen werden. Diese Zellen reagieren sehr empfindlich auf Umweltveränderungen und sich ändernde Kulturbedingungen. Das Einfrieren und Auftauen wird damit zu einem schwierigen Prozess für diese Zellen. Unseren Erfahrungen nach können wir einen Verlust von 30 % bis 40 % der lebenden Zellen nach jedem Einfrieren/Auftauen Zyklus nicht vermeiden. Zusätzlich benötigen die Zellen nach dem Einfrieren in unserem vorherigen Medium eines anderen Herstellers eine Erholzeit von mindestens fünf Tagen. Kürzlich haben wir das Bambanker Medium mit faszinierenden Resultaten getestet: Erholzeiten von nur zwei Tagen von 80 % der lebenden und "gesunden" Zellen. Wir konnten diese Verbesserung der Erholzeit und die Proportion von lebenden Zellen bei allen vier Zelllinien, die wir bisher getestet haben, beobachten. Wir sind sehr zufrieden mit Bambanker und werden es weiterhin benutzen.

on 10/24/2017

“JCRB cell bank has carried out cell bank business for 30 years and we currently store 1400 types of cell lines. In 2013, we offered about 4,300 ampules for a fee to domestic and foreign researchers. Due to the high number and the wide variety of cell lines, we had some problems. Thus some users complained that their cell lines of dying after thawing, resulting in unsuccessful cultivation. Especially four types of cell lines were a problem which had to be urgently improved. Therefore, we compared Bambanker™ with our currently used commercial preservation medium in a cryopreservation test. The cell lines, which were stored with Bambanker™, showed much higher cell proliferation than cells, which were stored with our currently used commercially available product. Surprisingly, with Bambanker™ we got for all four cell lines very reproducible results. In addition, Bambanker has no differences between the lots since it does not contain serum. We are very grateful for that because it strongly simplifies the delivery procedure overseas. In the future, we will completely change to Bambanker™ in order to improve the survival rate and growth of our cells. We are thankful for resolving that long-standing problem and recommend Bambanker™ to all domestic researchers and foreign cell banks. This data will be also described at our JCRB website. In the near future we plan to test Bambanker™ for human tissue-derived and human iPS cell lines.”
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