Frequently asked questions & protocols
Product Sheets->
Protocols
Landing pad stable cell lines
Frequently Asked Questions
How are the cells generated?
We use our landing pad technology to generate stable cell lines.
Each of our master cell lines feature a landing pad located at a distinct genomic locus. These loci are all chosen for their capacity to support stable expression of a diverse range of targets.
We then use a recombinase for directional DNA integration, essentially cutting out a part of the landing pad and inserting your target in place. Both, the recombinase and the target are delivered by co-transfection using plasmids.
For a detailed discussion of our landing pad technology, schedule an introductory call, or have a look at our methodology paper: Schucht et al. J Biomol Screen (2011)..
Can I use your stable cell lines for production of biologics?
While our landing pad cell lines are optimized for high and stable expression, the single-copy integration approach leads to lower titers compared to multi-copy approaches making our landing pad cells best suited for use in drug discovery.
Please also note that the cell culture conditions and protocols are not optimized for production, for example in fed-batch processes.
For which cell types do you offer landing pad technology?
Currently we have established our landing pad technology in:
- CHO
- HEK293
- Immortalized Llama cells
Please contact us, if you would like us to establish our landing pad technology in your preferred cell line.
We also offer the generation of stable cell lines in any cell background using viral and non-viral random integration.
Can you establish landing pads in other cell lines as well?
Yes, we also offer the establishment of our landing pads in other cell backgrounds as a service. Please contact us to discuss your project idea.
We also offer the generation of stable cell lines in any cell background using viral and non-viral random integration.
What cell line package do I need, Basic or Standard?
The different packages provide a framework for our projects and can be further customized even during an ongoing project. We will provide you with a package recommendation during our feasibility assessment.
Generally, we recommend the Basic package if you just need a cell line and the target is low-risk meaning it has no know toxicity and has already been successfully overexpressed in stable recombinant cells – either by us or by others.
With the Standard package, we will use multiple expression setups (combination of master cell line and promoter) to find the ideal combination for your target. This increases the chances of generating a cell line even for difficult-to-express targets and opens up the possibility to obtain sets of cell lines with different expression levels of the target.
Llama...??
That’s right. We have previously immortalized a Llama cell line, also known as CellVacc-Llama. It is a fibroblastoid cell line derived from a male Llama. We then established our landing pad technology in this cell line.
This is especially useful for cell-based immunizations as you can very quickly generate large numbers of stable recombinant llama cells overexpression your immunogen for immunization campaigns. Ideal for complex targets, such as T-cell receptors that need a membrane-bound configuration for stability.
The llama cell background minimizes background epitopes in cell-based immunizations compared to a xeno cell line such as CHO or HEK293.
Can I use the landing pad cells for commercial purposes?
Generally, most commercial research applications are allowed and you receive a royalty-free license to our IP to use the cells for your research with completion of the service project.
Any other commercial uses may be possible as well (offering research services, release assays, incorporation into your products) but they may require separate licenses.
What is the BSL of the stable cell lines?
The cell lines are BSL1.
What are the lead times for your stable cell services?
We do not have any lead times and can start your project immediately.
How do I order your cell services?
The first step is to get in touch with us and tell us about your project idea. We will then provide you with a quote, which you can use for generating a purchase order.
You can then submit this PO to and we will start the project.
If we need to be registered as a supplier in your database, please contact .
In addition, we are listed on multiple digital purchasing platforms.
Random integration stable cells
Frequently Asked Questions
How are the cells generated?
To generate cell lines in any cell line we use traditional random integration technology, with the option to generate either polyclonal pools or monoclonal cell lines.
We us a range of standard gene delivery systems; other technologies are available upon request.
Non-viral:
- Lipofection (various systems)
- Electroporation/Nucleofection
- Calcium phosphate
Viral:
- Lentivirus
Can I use your stable cells for commercial purposes?
Generally, most commercial research applications are allowed and you receive a royalty-free license to our IP to use the cells for your research with completion of the service project.
Any other commercial uses may be possible as well (offering research services, release assays, incorporation into your products) but they may require separate licenses.
What is the BSL of the stable cell lines?
If the parental cells are BSL1, the stable cell lines will be classified BSL1 as well.
What are the lead times for your stable cell services?
We do not have any lead times and can start your project immediately.
How do I order your cell services?
The first step is to get in touch with us and tell us about your project idea. We will then provide you with a quote, which you can use for generating a purchase order.
You can then submit this PO to and we will start the project.
If we need to be registered as a supplier in your database, please contact .
In addition, we are listed on multiple digital purchasing platforms.
What is the cost of a random integration stable cell line generation project?
Cost for stable cell line generation can vary significantly across different projects. Here are two example projects and estimated costs for each of them:
Polyclonal overexpression in standard cell line
Overexpression of a low-risk target in a standard cell line with established transfection protocols. Detection of the target by flow cytometry using a commercially available antibody. Generation of polyclonal pools only and banking of 15 Mio cells.
Estimated costs: 3 000–4 000€
Monoclonal reporter cell line in a non-standard cell line
Design of a reporter construct. Optimization of transfection protocols. Integration and validation of the reporter construct using 3 reference compounds. Assay establishment and optimization. Generation of dose-response curves as part of the cell line characterization. Generation of monoclonal cells and banking of a 150 Mio cell research cell bank.
Estimated costs: 15 000–25 000€
Please note that these examples are provided for reference only and pricing of your project may vary.
Immortalization
Frequently Asked Questions
How are the cells immortalized?
We use a custom library of immortalization genes encoded in lentiviruses for cell-type-specific immortalization. The simultaneous expression of multiple different immortalization genese stimulates different proliferation pathways meaning less overall expression is required to achieve robust proliferation. This leads to a better phenotype compared to traditional one- or two-gene immortalization approaches.
For a detailed discussion of our immortalization technology, schedule an introductory call, or have a look at our methodology paper: Lipps et al. Nat Comm (2018).
What is the BSL of the immortalized cell lines?
The immortalized cell lines are BSL1. Now, there may be exceptions to this, if the source material is already classified BSL2. For example some porcine material is considered BSL2 in certain countries.We are happy to assist you with determining the BSL of your material.
Will I learn the integrated immortalization genes?
Yes, we will provide you with a list of all integrated immortalization genes.
What are the cell types that you can immortalize?
Generally, any primary cell that:
- can be kept viable in culture for about a week
- grows adherent
- is derived from a mammalian species
is considered a low- to mid-risk project by us with good to very good chances of creating a functional cell line.
Here are some cell types and species that we consider mid- to high-risk projects and that likely require preliminary experiments:
- cells derived from the haematopoietic system
- cells from the nervous system (central and peripheral)
- cells from avian species
- cells from aquatic species
Can I use the immortalized cells for commercial purposes?
Generally, most commercial research applications are allowed and you receive a royalty-free license to our IP to use the cells for your research with completion of the service project.
Any other commercial uses may be possible as well (offering research services, release assays, incorporation into your products) but they may require separate licenses.
Please also note that this also depends on the source material, its ownership, any use limitations or exclusions as listed in the patient consent.
For how long will the cells proliferate?
We commit to providing you truly immortalized cells meaning they can be expanded indefinitely. However, as with any other cell line, regular monitoring of the phenotype (markers, assays) is recommended, especially once the cells are expanded beyond passage 50.
With proper cell banking practice, you should never run out of high-quality, low passage immortalized cells.
How many cells are needed for immortalization?
While anything above 1Mio cells is ideal, we can work with as low as 0.1Mio cells.
The more, the better, especially if the project involves characterization of the primary cells prior to immortalization or comparison of the immortalized cells with the primary cells.
What are the lead times for your immortalization services?
We do not have any lead times and can start your project immediately. Please note, however, that sourcing primary material can sometimes mean waiting times of several months especially for tissue sourcing.
How do I order your cell services?
The first step is to get in touch with us and tell us about your project idea. We will then provide you with a quote, which you can use for generating a purchase order.
You can then submit this PO to and we will start the project.
If we need to be registered as a supplier in your database, please contact .
In addition, we are listed on multiple digital purchasing platforms.
Protocols
CI-huArlo
– Human Alveolar Epithelial cells
-> View PDF
CI-huBroBEC
– Human Bronchial Epithelial cells
-> View PDF
CI-huEGC
– Human Enteric Glial cells
-> View PDF
CI-muHepa
– Murine Hepatocytes
-> View PDF
CI-pNAEC
– Porcine Nasal Epithelial cells
-> View PDF
CI-muMEC
– Murine Microvascular Endothelial cells
-> View PDF
CI-hAELVi
– Human Alveolar Epithelial cells
-> View PDF
CI-huFIB
– Human Fibroblasts
-> View PDF
CI-huMEC
– Human Microvascular Endothelial cells
-> View PDF
CI-huOB
– Human Osteoblasts
-> View PDF
CI-huVEC
– Human Umbilical Vein Endothelial cells
-> View PDF
CI-huThyrEC
– Human Thyroid Epithelial cells
-> View PDF
CI-muADINTESTI
– Murine Adult Intestinal Epithelial cells
-> View PDF
CI-muINTEPI
– Murine Intestinal Epithelial cells
-> View PDF
CI-muMEC
– Murine Microvascular Endothelial cells
-> View PDF