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Cell and Gene Therapy Core - Services 2023

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


Consultation & training

iPS services (e.g. reprogramming & gene-editing )

CRISPR (design & trouble-shooting)

Vector production (AAV & LV) & DNA scale-up


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 650SEK/h. 


iPS services

from patient to edited iPS

iPS reprogramming

Fibroblast derivation (4 000SEK)

  • Derivation, expansion, and freezing of fibroblasts from freshly collected skin biopsy,
  • Delivery of mycoplasma tested fibroblasts, at least 300k/vial x 3 vials.

mRNA-based reprogramming from human fibroblast cells (40 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 (3 x 300k cells).
  • quality controls as follows;  Sendai virus clearance, cell authentication (STR), pluripotency and undifferentiated state confirmation, molecular karyotype and iPS growth pattern analysis
  • Optional g-banding Karyotype (4000SEK/clone) 
  • Optional cell stocks (24 or 48 extra QC:ed stocks, 5 000SEK and 7 000SEK ), recommended 

Sendai based reprogramming from human fibroblast cells/blood (45 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 (3 x 300k cells) with quality controls as follows;  Sendai virus clearance, cell authentication (STR), pluripotency and undifferentiated state confirmation, molecular karyotype and iPS growth pattern analysis
  • Optional g-banding Karyotype (4 000SEK/clone) 
  • Optional cell stocks (24 or 48 extra QC:ed stocks, 5000SEK and 7000SEK), recommended 

CRISPR edits in hiPS

  • Edits can be performed in researcher- or core-provided iPS-lines
  • We offer
    • KO edits (30 000 SEK)
    • point mutation correction/insertions (38 000SEK)
    • reporter and tagged lines (38 000SEK + plasmid/template cost)
  • Design of the gRNAs and repair templates are included.
  • Edits will be performed using RNP nucleofection. 
  • Expected delivery of 2 correctly edited clones, (300k/vial x 5 vials) with basic characterization, including cell authentication, undifferentiated state (immunostaining) and molecular karyotyping.
  • Optional deliveries (price per clone)
    • cell stocks (22 extra vials, 3 500SEK), recommended 
    • additional edited clones
    • unedited clones
  • Optional QC of
    • g-banded karyotyping (4 000 SEK)
    • pluripotency test (tri-lineage differentiation, 4 000SEK)
    • Top 5 off-target analysis (2 500 SEK)

QC:ed iPS Cell Stocks

Coming soon

 

 

Access to control iPS lines

Core-generated control iPSC lines that can be distributed for user experiments. (3000SEK/2 vials)

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 (see above)
  • Experimental design (in a multitude of cell types and species) 650SEK/h
  • gRNA design (CRISPR-cut, CRISPRi, CRISPRa) 650SEK/h
  • HDR design and strategies for tagging and correction 650SEK/h
  • Cloning protocols for gRNA insertion
  • Access to off-the shelf plasmid backbones (15ug) 200SEK
  • Initial validation of CRISPRi and CRISPRa in HEK or iPSC (new, 6 500SEK)
  • Cut and edit validations strategies

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

 


Vector production services

Full service production of lentivirus and AAV.

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 vector preparations

How we produce the vector preps: Transfection of the packaging cell with the user provided transfer vector along with core provided packaging constructs, followed by ultracentrifugation/concentration of the supernatants. The concentrated supernatants are titrated via FACS or qPCR. 

Lentiviral vectors are offered in 3 sizes

  • mini, 2 000 SEK, delivers around 90ul of 107-108 TUI/ml*
  • standard, 4 300 SEK, delivers around 300ul of 107-108 TUI/ml*
  • large 11 500 SEK, delivers 1000ul of 107-108 TUI/ml* 

*depending on plasmid size and titration protocol

Useful Information:

  • For Lentiviral vectors please provide the following amounts of donor plasmid DNA: LV-mini 20ug, LV-standard 55ug, LV-large 160ug.
  • 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): 11 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
  • Robust quality control and titration of vectors
  • A wide variety of AAV serotypes (AAV 2, 5, 6, 8, 9)

Useful Information:

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

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


 


 

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Control considerations
Control considerations for CRISPR edits
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