Helsinki Virus (HelVi) Cores

 

AAV Gene Transfer and Cell Therapy Core Facility

The Helsinki Virus (HelVi) Core is a service platform that provides generation of recombinant adeno-associated viral (rAAV) preps for academic and industrial customers in Finland as well as abroad. HelVi uses biosafety level I laboratory equipped with all necessary cell culture equipment such as laminar hoods, incubators, centrifuges and microscopes.

Contact persons: Andrey Anisimov, +3582941 25562, andrey.anisimov@helsinki.fiTanja Laakkonen, +3582841 25563, tanja.laakkonen@helsinki.fiKari Alitalo, Core director, +3582941 25511, kari.alitalo@helsinki.fi

Location: Biomedicum Helsinki, 5th floor, Haartmaninkatu 8, Meilahti Campus

Brief introduction to the technology - 5 easy steps

  1. The gene-of-interest or shRNA are cloned into the AAV plasmid vector
  2. Generation of rAAV particles carrying the gene of interest or shRNA
  3. Administration of rAAV particles into cells or experimental animals
  4. rAAV transduces host cells and releases its recombinant genome
  5. The transgenic protein or shRNA is expressed

Services

  • Manufacturing rAAV preps from the customer’s gene-encoding plasmids. The available capacity allows the production of 3xE+11 to 3xE+12 virus genomes/ml in one purification round. The currently used serotypes are 2, 6, 8 and 9
  • Routine functional tests of the rAAV preps to characterize transduction efficiency and the ability to produce the transgenic protein in transduced host cells
  • Providing customers with control rAAV preps, immediately available upon request
  • Consulting the customers regarding rAAV vector design, tissue/cell specificity of transduction, fluorescent markers, tags, methods to increase expression level and bioavailability
  • Researcher training tailored to client needs
  • Generation of CRISPR rAAV preps for genome editing

Instruments

  • Heracell 150 CO2 cell culture incubator (2 x)
  • KOJAIR cell culture hood for the viral work
  • Sorvall RC 6+ centrifuge equipped with rotors
  • A&D EK-400H scale
  • Ultracentrifuge Optima XL-80K Beckman Coulter
  • EVOS ® FL Imaging System
  • Olympus CK2 inverted bright field microscope
  • Zeiss AxioImager and Zeiss LSM780 and 880 microscopes

Applications

  • Recombinant virus production and quantification
  • Quality test for the ready viral preps
  • Optimization of AAV vectors upon request

User fees

The research infrastructure is supported by University of Helsinki (HiLIFE and Research Programs Unit) and Biocenter Finland. The prices for University of Helsinki customers (VAT 0 %) are listed below. For the other customers, the VAT is 24%.

Service Fee
Internal customers 676,86 €
HUS customers 676,86 €
External academic customers 1050,27 €
External customers 1397,43 €

Ready AAV preps available upon request

HelVi has several ready AAV preps in storage, which can be mainly used as a negative control in vivo or in vitro. These ready AAV preps * (as listed in the Table below) can be sold in smaller amounts, reducing the price.

Transgene AAV serotype Notes
Human Serum Albumin (HSA) AAV9  
Empty vector (no transgene) AAV9  
Fc-part of mouse IgG (mFC) AAV9, AAV2 Control for Fc-fuced proteins
EGFP AAV9, AAV2, AAV8, AAV6  
Luciferaasi AAV9, AAV2, AAV8  
LacZ AAV9  
Cre recombinase AAV9, AAV2, AAV8 Can be used to cut away loxP flanked DNA sequences

* For other available AAV preps, not included in the Table, please, send e-mail or call.

BIO­MEDICUM VIRUS CORE - BVC

The Biomedicum Virus Core (BVC)

BVC is a service platform that provides biosafety level II laboratory space for recombinant virus production as well as consultation, technical help and gene transfer products for the researchers on the Meilahti and Viikki Campuses. BVC is equipped with basic cell culture equipment such as laminars, incubators, centrifuges and microscopes. Latest purchase for BVC is a new fluorescent microscope. All registered users are allowed to use the EVOS® FL Imaging System.

In order to work in BVC facility, you need to have a permission that is obtained through registration. If you want to start working in the facility, you need to take the BVC biosafety training (100€/person for UH customers) and be registered as a BVC user (use costs 19 €/h for UH customers including all BVC materials, reagents, waste management, equipment maintenance and partial support). If you are part of STEMM program, you can choose whether you want to be registered a BVC user or a STEMM user (no hourly fee, but you have to bring your own materials and reagents etc.) New STEMM users also need to take the BVC biosafety training. Old BVC users can continue working in the new facility as in the old one, but are advised to contact FuGU-support if they have any questions, if they want to become STEMM users or want to get a tour in the new facility. EVOS microscope users need to be registered and trained as well.

If you are not registered as a user and would like to get the Biosafety training or quick training to be able to use the EVOS microscope, please contact fugu-support@helsinki.fi, tel. 02941 25494.

Here you can find our updated Biosafety manual (2019)

In case of an emergency, please read BVC's rescue plan (in Finnish only) 

After registration working hours can be reserved through BVC online calendar

Contact person: Katariina Karjalainen (Tiina Raatikainen on maternity leave), BVC Management, +3582941 25494, fugu-support@helsinki.fi

Location: Biomedicum Helsinki, 5th floor, Haartmaninkatu 8, Meilahti Campus

Services

  • Registration & Biosafety Training
  • General facilities supervision and support
  • Hood and facility use
  • Biosafety test (RCV test)
  • Virus titering test for lentiviral particles (p24 test)
  • General cell lines related to virus work
  • Transfection reagent aliquotes

Instruments

  • Thermo Forma Series II Water Jacketed CO2 Incubator (2 x)
  • Heraeus Multifuge 3S-R (w/ tube and plate rotor)
  • Olympus CK40 bright field microscope
  • A&D EK-400H scale
  • Labnet International Spectrafuge 24 D
  • Wealtec E-centrifuge
  • Hettich Universal 32 centrifuge
  • Memmert waterbath
  • Sartorius Biohit MidiPlus pipettors
  • Stuart vortex
  • EVOS® FL Imaging System

Applications

  • Recombinant virus production
  • Transduction of target cells
  • Fluorescent wide-field imaging for BSL2 substances

RCV & p24 tests

RCV Test? Recombinant retro- and lentiviruses are designed to perform only one round of infection. After the transduction, a segment of the transfer vector that encodes shRNA/miRNA or cDNA integrates into the host cell's genome and is expressed under the control of the viral promoter. The recombinant viruses do not carry genetic information for production of virus particles and, therefore, cannot replicate or produce new virus particles in the target cells. However, there is a very small likelihood that unusual recombination events lead to formation of replication competent viruses during the virus production. We exclude the presence of replication competent virus (RCV test) in lentivirus preparations upon request.

RCV test can also be performed to transduced cells to ensure that the cells are virus-free before transferring cells outside from the Biosafety Level 2 laboratory.

Capsid Titer Test (p24)? A 24 kilodalton protein (p24) is a major structural core component in the recombinant lentiviruses. We use a p24 specific enzyme-linked immunosorbent assay (ELISA) to measure the p24 lentivirus capsid protein concentration (pg/ml) from the cell culture supernatants containing lentivirus preparations. 

How does it work? FuGU provides RCV and p24 titer tests as a service. These tests are performed every other Wednesday. The test dates can be found from the BVC Online Hood recervation calendar. For RCV test we need 1 ml unfiltered cell culture media (at least 24 hours after splitting cells or media change) and for p24 titer analysis we need 10 μl of viral supernatant. Label samples with your initials and running numbers (KK1, KK2..). If you are a registered BVC facility user, you can bring your samples into the -20C sample box at the BVC facility. If you are not, please contact us to arrange the sample delivery. Also remember to send an e-mail to fugu-support@helsinki.fi containing your sample information and a billing reference in order to have your samples analyzed!

User fees

The research infrastructure is supported by University of Helsinki (HiLIFE and Research Programs Unit) and Biocenter Finland. The prices for University of Helsinki customers (VAT 0 %) are listed below. Other customers, please contact fugu-support@helsinki.fi.

Service Price (eur)

Registration & Biosafety training (€/person)

100

Usage of the facility/h (other than STEMM progam users)

19
RCV test 45
p24 test 55
Cell ampoul 10
Transfection reagent 200
BVC services