Purpose
The primary purpose of the Semmelweis University Cell Core Facility is to help all researchers at the university providing stable, cost-effective, and safe virus transduced cell lines. New, stable cell lines can facilitate research towards new directions and open new possibilities.
Background
Compared to transient expression, stable cell lines incorporate the target DNA sequence into the genom, allowing long-term, stable gene expression. These features make these cell lines reliable on long-term and make the experiments reproducible.
Aims
Our aim is to generate stable cell lines using lentiviral vectors for research groups that either lack the facilities to work with viruses or require our expertise in this area. We can produce cell lines to support gene overexpression (cDNA, CRISPR activation), gene knockdown (shRNA, CRISPR interference), or custom gene editing using the CRISPR/Cas9 system. If you have other genome editing plans, please contact us to discuss the possibilities for custom-designed cell lines.
Workplan
Researchers provide the plasmid of interest and the desired cell line to our lab. Vector plasmid design can also be arranged through consultation with our team. The Core Facility will assist in selecting the appropriate promoter, CRISPR strategy, and vector design tailored for either in vivo or in vitro applications.
Following design and cloning (if required), viral vector production begins and typically takes 4 to 6 weeks. After titration of the lentiviral stock by RT-qPCR, we perform an MOI (multiplicity of infection) test to determine the optimal virus concentration for the specific cell type.
Using the established MOI, we transduce the cells. Stable cell lines are then generated via FACS (fluorescence-activated cell sorting) or antibiotic selection. Monoclonal cell lines can also be produced through single-cell sorting.
Before delivery, all cell lines undergo Mycoplasma testing. We provide the final stable, non-infectious cell lines for downstream validation, and we can assist with expression verification using RT-qPCR upon request.
Services
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Consultation on construct design and cloning strategies
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Full construct building and cloning service
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Construct testing and validation (e.g., restriction digestion, sequencing)
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DNA scale-up (Maxiprep or other preparation methods)
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Full-service lentivirus production:
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Transfection of vector plasmid into HEK 293T cells
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Collection and concentration of viral supernatant via ultracentrifugation
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RT-qPCR-based viral titration
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MOI (multiplicity of infection) testing on the target cell line
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Cell handling, Mycoplasma testing, expansion, and transduction
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Antibiotic selection or FACS sorting for stable cell line generation
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Cryopreservation of Mycoplasma-tested cells after one week of expansion
Charges
The Cell Core Facility operates on a non-profit basis within Semmelweis University. All fees cover the cost of consumables and personnel time.
Our services are available to both internal and external academic users. Semmelweis University researchers have priority access. For external academic or industrial customers, please contact us directly for pricing and service availability.
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Initial consultation and planning are free of charge.
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Further consultation and design: 4,000 HUF/hour
A typical stable cell line production costs approximately 500,000 HUF, depending on the specific services required.
The final price is based on the actual combination of selected services:
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Full planning and construct design: included in initial estimate
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Molecular biology services:
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MAXIPREP plasmid purification: 50,000 HUF
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Construct building / full cloning: 150,000 – 250,000 HUF (depending on complexity)
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Lentivirus production: 150,000 HUF per batch
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FACS sorting: 20,000 HUF/hour
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Cell line handling (including expansion, selection, and validation):
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First cell line: 280,000 HUF
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Additional cell lines: 50% of the full price
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RT-qPCR expression validation: 75,000 HUF
All prices include taxes and university overheads.
To request services or discuss your project, please contact us by email or phone.
Contact
Ágnes Varga, PhD
Phone: +36 1 459 1500 / 60369
Mobile: +36 30 016 4787
E-mail: varga.agnes1@semmelweis.hu
0.112 laboratory room
D part, ground floor
Basic Medical Science Center
Institute of Translational Medicine
Semmelweis University
1094 Budapest, Tűzoltó street. 37–47