With years of experiences with cell line cell immortalization, scientists at abm
have been able to successfully immortalize essentially any human and mouse cell in last 10 years using different immortalizing agents or genes. If you have a primary cell culture that needs to be immortalized, our custom services is the quick and reliable service for your project. It saves you time and money over trying different genes yourself. Our custom service package already includes all different genes and you will not pay anything unless your cells are successfully immortalized. So it is risk-free and you pay only when your cell line is immortalized.
We not only provide cell immortalization custom service, but also sell cell immortalization products.
Contact us for quotation of our cell immortalization services.
Tel: 1-866-757-2414 (Toll Free) , 604-247-2416 (Local)
How do I send my cells and place an order? and What will I be receiving?
For frozen cells, you must ship in ample dry ice and each vial must contain 1-2 million cells. Another method is to send us live cells. If the cells are fast growing, we recommend sending three T25 flasks at 70% density. If the cells are slow growing, we recommend sending two T25 flasks at 90% density. The flask should be filled with complete medium without any air bubble and at room temperature. The flask can be filled with medium using a 50ml culture tube. Make sure that sterilization procedure is strictly observed. For North American customers, the cells should be sent on Monday. For non-North American customers, the cells should be sent on Friday or Monday to avoid transit delay over the weekend. Please note that the cells are immortalized in the condition that they were received and we cannot be liable for any changes during transit. We ask that customers supply a picture of your primary cells growing in culture to depict 'original' cell morphology when setting up shipment. This is to help us assess the cells we receive are in good condition before we start the immortalization project.
If your cells require medium other than DMEM and RPMI, we will also require 1L of the specified medium and any applicable growth factors or supplements to be provided by you for the project. Please submit all components of the complete media (e.g. if growth factors, cytokines etc. are applicable) individually to eliminate potential degradation of components in the media during transit. Kindly include instructions for making the complete media in your shipment. Additionally, please provide at least 5 x coated 6-well plates and 10 x T25 flasks if the cells require specially treated culture vessels. Once you have shipped your cells, please forward us the tracking number for custom clearance. Information on how to ship cell samples to abm can be found on our support page: https://www.abmgood.com/Technical-Support.html
Once we receive the cells, we will performed custom cell immortalization in 6-well plates with all our cell immortalization reagents, including but not limited to, SV40 T antigen, hTERT, Myc, E6/E7.
We will make sure the cells are immortalized and have the ability to grow forever. The final product to be shipped to you with be two to three different clones that have been successfully immortalized.
Can I purchase immortalized human neurons? I don't have primary neurons, but rather you can provide both the primary cell and the immortalization service?
Yes, we will be able to try to immortalize those cells for you. Please let us know your specific requirement and we will send you an official quotation.
How many cells do I need to send in order to immortalize one cell line?
For frozen cells, you must ship in ample dry ice and each vial must contain 1-2 million cells. Another method is to send us live cells. If the cells are fast growing, we recommend sending three T25 flasks at 70% density. If the cells are slow growing, we recommend sending two T25 flasks at 90% density.
What should I do if my cells do not grow using standard media?
If your cells do not grow using DMEM + 10% FBS or RPMI + 10% FBS, then we need at least 1L of your growth media plus additional amounts of associated growth factors.
What kinds of methods are you going to take to demonstrate the lentivirus-infected cells are immortalized and whether the cells characteristics are changed after immortalization?
Usually if the cells show positive transgene expression in qPCR and are passaging well in comparison to the primary cells, that is a confirmation of immortalization. In one of the rare events, if the trangene expression is lost over some late passages, we may need to re-immortalize.
Where applicable, the cells will be selected by antibiotics then subsequently passaged 30+ beyond the normal stage of senescence to confirm immortalization. We closely monitor typical cell behavior including growth rate and morphology., however all downstream characterization must be determined by the end user. We do not conduct gene expression assays as part of this service. Please note there may be sometimes changes in any markers or morphology due to immortalization and we do not claim that all or any specific markers be retained after immortalization.
Does the service perform western blot analysis or qPCR analysis in order to confirm the cells were successfully immortalized?
qPCR analysis is performed on the transgene expression
Please let us know details of the deliverables from this service.
We provide up-to 2 frozen vials of the best clones as a result of testing individual reagents. If multiple genes succeed in immortalizing the cells, we can provide 2 vials for each successful reagent depending on the cell growth, upon request (additional charges may apply depending on scope of request).
QC data provided in a final custom service report will include:
MORPHOLOGY ASSESSMENT, QPCR ANALYSIS ON THE TRANSGENE EXPRESSION, TEST RESULTS FOR PRESENCE OF MICROBIAL CONTAMINANTS