The browser you are using is not supported by this website. All versions of Internet Explorer are no longer supported, either by us or Microsoft (read more here: https://www.microsoft.com/en-us/microsoft-365/windows/end-of-ie-support).

Please use a modern browser to fully experience our website, such as the newest versions of Edge, Chrome, Firefox or Safari etc.

Cell and Gene Therapy Core - Services

Services are available to all internal and external academic customers (international customers please contact us for pricing). StemTherapy and Multipark users have priority access. 

Consultation/trainingiPS, CRISPR, Vector production, Cloning


Consultation and training

We offer training, consultation, design and trouble-shooting in all core related areas. 

First meeting is always free, further consultation, training and designs are charged at 250SEK/h. 


iPS services

 

iPS reprogramming

Fibroblast derivation (3 000SEK)

  • Derivation, expansion, and freezing of fibroblasts from freshly collected skin biopsy,
  • Delivery of mycoplasma tested fibroblasts, at least 500k/vial x 3 vials.

mRNA-based reprogramming from human fibroblast cells (25 000SEK)

  • Reprogramming is done with feeder free system.
  • Picking at least 12 colonies
  • Expansion of top three to four colonies
  • Deliver 2 clonally expanded mycoplasma free iPSC lines  at p5-p7(at least 500k/vial x 3vials) with basic characterization, including cell authentication, immunostaining (Oct3/4, Sox2,Nanog,Tra160, Tra181).

Sendai based reprogramming from human fibroblast cells/blood (30 000SEK)

  • Reprogramming is done with feeder free system
  • Picking at least 12 colonies
  • Expansion of top three to four colonies
  • Deliver 2 clonally expanded mycoplasma free iPSC lines at around p7 (at least 500k/vial x 3vials) with basic characterization, including cell authentication, immunostaining (Oct3/4, Sox2,Nanog,Tra160, Tra181)
  • If needed, we can run test for presence of sendai virus, we charge 2000kr/clone/test every 3-5 passages after passage 10

CRISPR edits in hiPS

  • Edits can be performed in researcher- or core-provided iPS-lines
  • We offer
    • KO edits (20 000 SEK)
    • point mutation correction/insertions (20 000SEK)
    • reporter and tagged lines (25 000SEK)
  • Design the gRNAs and repair templates are included.
  • Edits will be performed using either RNP nucleofection or plasmid-based delivery. 
  • Expected delivery of 2-3 correctly edited clones, (500k/vial x 3vials) with basic characterization, including cell authentication, immunostaining (Oct3/4, Sox2,Nanog,Tra160, Tra181)
  • Optional deliveries
    • additional edited clones (2 000 SEK/clone)
    • unedited clones (2 000 SEK/clone)
  • Optional QC of
    • g-banded karyotyping (4500SEK/clone)
    • Top 5 off-target analysis (2000SEK/clone)

 

Access to control iPS and ES lines

Core-generated control iPSC lines that can be distributed for user experiments. (2000SEK/2 vials)

We have also banked numerous Harvard and WiCell generated human embryonic stem cell lines and have agreements in place to distribute these lines to users via an accelerated MTA approval contract at significantly reduced cost to local users. (1000SEK)

User access to Cell Culture Clean Rooms

The facility is complete with advanced and state-of-the-art pluripotent stem cell tissue culture equipment, reagents and cell storage facilities all maintained by the core facility staff. Users of the clean rooms must undergo a 1 hour introduction training prior access. LAF hoods are booked via an online calendar. Users may use the facility in a limited and availability dependent manner. The work in the core is limited to ES and iPS cell culture, maintenance, and expansion; not differentiation of the pluripotent stem cell lines. 


CRISPR

  • CRISPR edits in iPSCs (new, see above)
  • Experimental design (in a multitude of cell types and species) 250SEK/h
  • gRNA design (CRISPR-cut, CRISPRi, CRISPRa) 250SEK/h
  • HDR design and strategies for tagging and correction 250SEK/h
  • Cloning protocols for gRNA insertion
  • Access to off-the shelf plasmid backbones (15ug) 200SEK
  • Cloning & vector production services (see below)
  • Initial validation of CRISPRi and CRISPRa in HEK or iPSC (new, 6 500SEK)
  • Cut and edit validations strategies
    • Pipeline for NGS amplicon sequencing
    • Referral to TigerQ  (complete service)

cellandgenetherapy [at] med [dot] lu [dot] se (Contact us) for CRISPR designs and edits.

 


Vector production services

Full service production of lentivirus, oncoretroviral and AAV.

Our staff has a long experience in the production of viral-based vectors and has become an important technological resource for researchers, providing services to non-profit institutions and academic investigators worldwide.

The core provides investigators access to state-of-the-art vector technology for preclinical studies and other basic research applications to be able to provide users with highly reproducible, high-titer vector preparations. 

 

Lentiviral or Oncoretroviral vector preparations

Our trained staff will take the vector plasmid you provide us and within 4 weeks you will receive a high titer vector preparation. We have the technical experience as well as state-of-the-art infrastructure

How we produce the vector preps:

1) transfect the packaging cell with the user provided transfer vector along with core provided packaging constructs

2) collect and filter vector containing supernatants

3) ultracentrifugation/concentration of the supernatants

4) titration of vector supernatant via FACS or PCR

5) vectors will be delivered in 10ul/tube or 50ul/tube aliquots (depending on selected concentration

Useful Information:

  • Lentiviral vectors are offered in 3 sizes
    • mini, 1 500 SEK
    • standard, 3 300 SEK
    • large 9 000 SEK
  • at two concentration level (normal and high). 
    • standard batch - high concentration, delivers 80-100ul of 108-109 TUI/ml*
    • standard batch - normal concentration, delivers 300ul of 107-108 TUI/ml*

              (*depending on plasmid size and titration protocol)

  • For Lentiviral vectors please provide the following amounts of donor plasmid DNA: LV-mini 15ug, LV-standard 45ug, LV-large 135ug.
  • For Retroviral vectors provide 30ug of donor plasmid DNA.
  • The quality of the DNA matters, please ensure that you have performed restriction digest and/or sequencing and have an OD ratio of 1.7-2.0 and deliver at a concentration of 0.5-1.0ug/ul. 

If you have donor plasmids ready then you can download our order form. 

cellandgenetherapy [at] med [dot] lu [dot] se (Contact us) for vector related services.

 

AAV vector preparations

The AAV services provided by the Cell and Gene Therapy Core core are:

  • AAV based vector production (200ul, approx. 1·1014 - 1·1015 VG/ml): 8 000SEK
  • Storage and handling of core-produced AAV vectors according to agreement. 
  • Production of custom vectors designed according to the requests of the researcher
  • Consultation and advice in the design of custom vectors and in vector serotype selection
  • Design, cloning and production of plasmid DNA for the production of custom vectors (4 500SEK)
  • Robust quality control and titration of vectors
  • A wide variety of AAV serotypes (AAV 2, 5, 6, 8, 9)

Useful Information:

  • For AAVl vectors provide 100ng of the transfer vector plasmid, deliver at a concentration of 0.5-1.0ug/ul. 
  •  please ensure that you have performed with SmaI to verify ITR region.

If you have donor plasmids ready then you can download our order form. 

cellandgenetherapy [at] med [dot] lu [dot] se (Contact us) for vector related services.


Cloning

  • Construct design and cloning strategies consultation
  • Construct building / full cloning service (4 500 SEK)
  • Contruct testing and validation e.g. restriction digestion, sequencing
  • Maxiprep Service (or other DNA scale-up) (2 000SEK)
  • Related molecular biology services (tell us about your molecular biology needs) 

cellandgenetherapy [at] med [dot] lu [dot] se (Contact us) for cloning related services.


 

cell and gene
 
reprogramming
white box
Control considerations
Control considerations for CRISPR edits
blank
blank
 
 
beata in the lab