Archive for the 'Q&A' Category

Published by kawaguchi on 03 4月 2023

1-14 Level of physical containment of facilities

JCRB Cell Bank is testing the cell lines for viruses pathogenic to humans as extensive as possible. However, there is the problem of detection limit and it is practically impossible to examine “all pathogens” as well as unidentified viruses.

Therefore the cell line should be handled as potentially biohazardous materials. Practically, the handling in accordance to biosafety level 2 is recommended. This does not mean that the cell line produces BSL-2 pathogens, but is needed to avoid potential risk.

Published by kawaguchi on 28 7月 2015

Q12. When does the claim expiration end?

JCRB accepts complaints within 3 month after shipment. First, check Q&A page for troubles in cell culturing. If you need help, please contact JCRB with “Contact” page.

Published by kawaguchi on 28 7月 2015

Q1-8. Unable to make any prepayments.

For customers in except the USA, Canada, the Republic of Korea, Australia, New Zealand, Oceania and Europe including the below countries:
Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, United Kingdom and Switzerland)

We accept advance payment by Credit Card or Bank Transfer. So if you can’t make prepayments, please select Credit Card payment, or place an order through a distributor/an agent that you already trade with.

Published by kawaguchi on 28 7月 2015

1-11. Can I share JCRB cell lines to other researchers?

In principle, transferring JCRB cell lines to a third party is not allowed.
When we receive a request for cells, applicants must sign the “Request and Agreement Form (Form A)”, which specifies that applicants should not transfer the cell lines to a third party. It is important to protect the intellectual rights of the depositor/developer of cell lines and also to assure the quality of JCRB Cell Lines. Please note that the above terms do not apply for collaborative research purposes.

However, because all cell lines have different limitations, please contact us if you are not sure if specific cell lines can be shared or not.

Published by kawaguchi on 28 7月 2015

1-10. How secure is my personal information when I order?

We take protection of your personal data very seriously, and utilise Secure Sockets Layer (SSL) software, which encrypts the information you input.

Published by kawaguchi on 28 7月 2015

1-12. About cell information

A quick keyword search is available on the ‘Cell Search’ page. Every ‘Cell Information’ page has a lot of useful information, with references. Although the data on each lot number including out of stock is available for information purposes, users cannot request a specific lot. If there is any other important information on a cell to be aware of, there will be an “Important Notice” icon at the top pf the page.

Published by kawaguchi on 28 7月 2015

1-13. What documents are required for importing cell lines?

We are based in Japan. The customs authority of your country may require you to obtain a special lisence or documents to import Human or Animal cell lines. Import fees may also apply.
For details, please inquire with authorities in your country.
In some case, we are unable to ship cell lines due to customs regulations in certain countries. Cell lines which are regulated by CITES (Wasington Trustee) are not available for users outside Japan.

Published by kawaguchi on 28 7月 2015

Successful cell culture

1. Cell culture process

Following the correct thawing procedure of frozen cells is very important for ensuring successful culturing.
  1. Do not allow the cell density in the dishes to become too dilute. It may restrict growth in the first culture.
  2. Determine viability when you open frozen vials. Especially in suspension cultures, it is difficult to separate dead cells and living cells.
  3. Check the mitotic rate of cell lines. Each cell line has a different mitotic rate, from about 12hr to 2 days or more.
  4. Follow to the instructions carefully. Differentiation and aging may cause a proliferation disorder.
  5. Change the culture medium at appropriate intervals. These depend on the cell lines and cell density. If the cell density is high, you should renew medium at shorter intervals. Likewise, keep long intervals if the cells are at low density.
  6. Subculture the cell lines at appropriate times. Post-confluent cells may undergo differenciation and exhibit reduced proliferation after passaging.

2. Guidelines for maintaining CO2 incubator

Keep the incubator clean and regularly check inside temperature, humidity and CO2 concentration. Ensure it is regularly serviced and calibrated, according to the manufacturer’s instructions.

3. Dishes and flasks

Please use appropriate cell culture dishes or flasks. Suspension culture grade or microbial grade dishes are inadequate for growing adherent cells. The size of dishes and flasks critically affects the survival and growth of cells at the first culture. In general, each vial of JCRB cell lines is prepared to start cell culture in one T-25 flask or one 6-cm dish. Do not use dishes or flasks that are too large or small, as further growth may be adversely affected.
Back to Q&A

Published by kawaguchi on 28 7月 2015

Just after receiving cells

1. What need to do when cell lines arrive?

  1. Frozen cells are cryopreserved in a vial and will be dispatched by courier in dry ice.
  2. It is recommended to start cultivation upon receipt of the cells, according to the optimum conditions for growth of that cell.
  3. If you with to store frozen vials, we ask you to transfer vials to the vapor phase of liquid nitrogen immediately after arrival. Do not immerse the vials in the liquid phase, as this could cause them to burst.
  4. Cells can be stored in a low temperature freezer at below -80°C for short-term storage of up to 30 days. Do not store them at -30°C, as this results in a rapid decrease in viability.

2. Can I store cell vials in liquid nitrogen?

Do not store vials in liquid nitrogen directly. If vials are immersed directly in liquid nitrogen (liquid phase), it may cause explosion of imperfectly sealed vials. It is recommended to reserve these vials in the vapor phase of liquid nitrogen tank.    

3. How do I start culturing and safely open glass ampules?

  1. Remove a vial from the package using safety gloves and a face shield to avoid injury from possible explosion.
  2. Thaw the vial(s) one by one within 2 min by shaking in moderately warm water (below 37°C).
  3. Under aseptic conditions, sterilize the entire surface of the vial with gauze moistened with 70% ethanol or cationic detergent sterilizer, then open the vial. If the vial is glass ampoule, cut the neck off while wrapped in sterilized gauze. All glass ampules are pre-scored around the neck to be easily cut off, but please take extra care to avoid injury.
  4. Transfer the cell suspension to a centrifuge tube, add 10 ml of the medium described in the cell data, centrifuge the mixture at 1000 rpm for 3 min, and discard the supernatant.
  5. Re-suspend the cells in the medium. Culture them in a flask or a dish using the standard method. The appropriate culture volume would be 5 ml per vial in a 25cm2 flask or 6cm dish, however, the volume of dishes must be flexible according to actual cell density. Make sure of cell proliferation before proceeding to passage, because excessive dilution may lead to cell death, especially in hemocytes and lymphocytes.
Back to Q&A

Published by kawaguchi on 28 7月 2015


1. Importatnt notes about media

  1. Do not use media that has been left in storage for a long time. Liquid media should be used within 3 months, powder media within 12 months after preparation.
  2. Keep media in good condition. Media deteriorates with repeated heating and cooling, or if it is stored under incorrect temperature.
  3. Serums, growth factors or other supplements must be added appropriately to cultures. Serums lot variation often affects to cell growth or concentration. Growth factors may suffer reduced efficiency due to incorrect handling.
  4. Check the pH of your medium when you make liquid medium from powder. You can judge pH by the changes in color of medium during cultivation.
  5. Does your medium include L-Glutamine or Sodium Bicarbonate?  L-glutamine is essential amino acid, but some commercially available media does not contain it. In this case, you must add L-glutamine to media separately during preparation. Also check whether they should be added before or after autoclaving.

2. May I use my own media instead of media in the cell data sheet?

The medium described in the cell data must be used to ensure the frozen cells will return to viability. Then, if required, please try to culture in your own medium. If you use a media other than that specified in the instructions, JCRB does not take responsibility for cells in case of a failure to grow, or other unforeseen circumstances.

3. Glucose concentration for DMEM medium

If the recommended basal medium is DMEM, please keep in mind that the glucose concentration in standard DMEM is 1 g/L. (low glucose DMEM)   Because high glucose DMEM is a modified form, you should use it only if it is specified in the cell line’s data.
Back to Q&A

Next »