Remove medium and rinse the adherent cells using PBS without calcium and magnesium (3-5 ml PBS for T25, 5-10 ml for T75 cell culture flasks). Add Accutase (1-2 ml per T25, 2.5 ml per T75 cell culture flask), the cell sheet must be covered completely. Incubate at ambient temperature for 8-10 minutes. Carefully resuspend the cells with medium (10 ml), centrifuge for 3 min at 300 g, resuspend cells in fresh medium and dispense into new flasks which contain fresh medium.
2 - 3 times per week
CM-1 (CLS catalog number 800150)
Handling of cryopreserved cultures
The cells come deep-frozen shipped on dry ice. Please make sure that the vial is still frozen. If immediate culturing is not intended, the cryovial must be stored below -150°C after arrival. If immediate culturing is intended, please follow the below instructions: Quickly thaw by rapid agitation in a 37 °C water bath within 40-60 seconds. The water bath should have clean water containing an antimicrobial agent. As soon as the sample has thawed, remove the cryovial from the water bath. A small ice clump should still remain and the vial should still be cold. From now on, all operations should be carried out under aseptic conditions. Transfer the cryovial to a sterile flow cabinet and wipe with 70% alcohol. Carefully open the vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of culture medium (room temperature). Resuspend the cells carefully. Centrifuge at 300 x g for 3 min and discard the supernatant. The centrifugation step may be omitted, but in this case the remains of the freeze medium have to be removed 24 hours later. Resuspend the cells carefully in 10 ml fresh cell culture medium and transfer them into two T25 cell culture flasks. All further steps are described in the Subculture section.
Handling of proliferating cultures
One or two cell culture flasks come filled with cell culture medium. Collect the entire medium in 1 or 2 x 50 ml centrifuge tubes, respectively. Carefully add 5 ml of cell culture medium to each T25 cell culture flask. Control the cell morphology and confluency under the microscope. Incubate at 37 °C for a minimum of 24 hours. Spin down the collected medium at 300 x g for 3 minutes to collect the cells which may have detached during transit. If a cell pellet is visible, resuspend the cells in 5 ml of cell culture medium and transfer to a T25 cell culture flask. Incubate at 37 °C for a minimum of 24 hours.
Quality control / Genetic profile / HLA
Mycoplasma contamination was excluded through PCR-based and luminescence-based mycoplasma assays. Bacterial or fungal contaminations are detected through daily visual cell monitoring.
References "HSC-T6 "
To identify mycoplasma contaminations we perform PCR-based and luminescence-based mycoplasma assays. We further determine any bacterial or fungal contamination through our standardized manufacturing processes.
Besides genomic DNA, RNA, cell pellets, and cell lysates in bulk, we can offer large quantities of assay-ready cells, plated cells in multiple formats, and frozen or growing cells. Contact us to receive a quote.
Each manufactured batch of cell lines* is authenticated via STR analysis. Contact us if you require a publication-ready STR report for your cells (*human, hamster, mouse, rat, and dog cells).
HLA characterization is available from more than 200 cell lines. HLA class I -A, B, C, and Class II HLA-DPA1, -DPB1, -DQA1, DQB1, and DRB1 alleles were obtained by next-generation sequencing methodologies (NGS) for class I and class II alleles.
This Modal is powered by moori Foundation
Functional cookies are absolutely necessary for the functionality of the web shop. These cookies assign a unique random ID to your browser so that your unhindered shopping experience can be guaranteed over several page views.
Information on origin:
The cookie stores the referrer and the first page visited by the user for further use.
Saves which cookies have already been accepted by the user for the first time.
The cookie is used to store the cookie settings of the site user over several browser sessions.
The CSRF token cookie contributes to your security. It strengthens the security of forms against unwanted hacker attacks.
The cookie is used to differentiate the cache for different scenarios and page users.
The cookie is used to provide the system with the user's current time zone.
The cookie is used by the payment provider to increase security when processing payments on the website.
The session cookie stores your shopping data over several page views and is therefore essential for your personal shopping experience.
Marketing cookies are used to display advertisements on the website in a targeted and individualized manner across multiple page views and browser sessions.
The cookie is used by Google AdSense to promote advertising efficiency on the website.
Google conversion tracking:
The Google Conversion Tracking Cookie is used to track conversions on the website effectively. This information is used by the site operator to target Google AdWords campaigns.
These cookies are used for statistics and shop performance metrics.
Conversion and usertracking via Google Tag Manager
Necessary for the appropriate function of our search solution.
Tracking cookies help the shop operator to collect and evaluate information about the behaviour of users on their website.
Google Analytics is used for traffic analysis of the website. Thereby statistics about website activities can be created and read out.