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Cell and Gene Therapy Core - Services 2023

***Happy New 2024***
New prices from the 1st of January.

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


Consultation & training

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

mRNA production

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 1000SEK/h. 


iPS services

from patient to edited iPS

iPS reprogramming

Fibroblast derivation

  • 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

  • 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; cell authentication (STR), pluripotency and undifferentiated state confirmation, molecular karyotype and iPS growth pattern analysis
  • Optional cell stocks, recommended 

Sendai based reprogramming from human fibroblast cells/blood

  • 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, karyotype and iPS growth pattern analysis
  • Optional cell stocks , recommended 

CRISPR edits in hiPS

  • Edits can be performed in researcher- or core-provided iPS-lines
  • We offer
    • KO edits
    • point mutation correction/insertions
    • reporter and tagged lines
  • 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 15 vials) 
  • QC included; basic characterization, including cell authentication, undifferentiated state (immunostaining/FACS) and molecular karyotyping.
  • Optional deliveries (price per clone)
    • extra cell stocks (24 or 48 extra vials)
    • additional edited or unedited clones
  • Optional QC of 
    • g-banded karyotyping
    • pluripotency test (tri-lineage differentiation)
    • Top 5 off-target analysis 

QC:ed iPS Cell Stocks - Master Cell Banks

Cellstocks overview

Cell stocks 20x300k cells, small QC panel, 

Cell stocks 48x300k cells, small QC panel, 

Cell stocks 20x300k cells, large QC panel, 

Cell stocks 48x300k cells, large QC panel, 

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. 

Hood booking: 300SEK/hour,  Reagents: List price. Introduction: 500SEK


 

mRNA production

validated mCherry or GFP mRNA; aliquots of 1,5 pmol  (1,5 ul)      3000 SEK

Custom mRNA production;                                                                 15000 SEK (plus plasmid cost)

 


CRISPR

CRISPR overview
  • CRISPR edits in iPSCs (see above)
  • Experimental design (in a multitude of cell types and species) 
  • gRNA design (CRISPR-cut, CRISPRi, CRISPRa) 
  • HDR design and strategies for tagging and correction 
  • Cloning protocols for gRNA insertion
  • Access to off-the shelf plasmid backbones (15ug) 
  • 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. 

vector preparation overview

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 **Please note new names of our sizes!!***

  • small (1 plate), delivers around 90ul of 107-108 TUI/ml*, 3000 SEK
  • medium (3 plates) , delivers around 300ul of 107-108 TUI/ml*, 6000 SEK
  • large (10 plates), delivers around 1000ul of 107-108 TUI/ml*, 15000 SEK

*depending on plasmid size and titration protocol

Useful Information:

  • For Lentiviral vectors please provide the following amounts of donor plasmid DNA: LV-small 20ug, LV-medium 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.

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. 1·1013, 12000SEK per batch)
  • Storage and handling of core-produced AAV vectors according to agreement. 
  • 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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