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

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 cell stocks (24 or 48 extra vials, 3 500SEK and 6 000SEK), recommended

Sendai based reprogramming from human fibroblast cells/blood (50 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, karyotype and iPS growth pattern analysis
Optional cell stocks (24 or 48 extra vials, 3 500SEK and 6 000SEK), 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 3 vials)
QC included; basic characterization, including cell authentication, undifferentiated state (immunostaining/FACS) and molecular karyotyping.
Optional deliveries (price per clone)
- cell stocks (24 or 48 extra vials, 3 500SEK and 6 000SEK), recommended
- additional edited or unedited clones (4 000 SEK/clone)
Optional QC of
- g-banded karyotyping (4 000 SEK/clone)
- pluripotency test (tri-lineage differentiation, 3 000SEK/clone)
- Top 5 off-target analysis (2 500 SEK/clone)

QC:ed iPS Cell Stocks NEW

Cell stocks 20x300k cells, small QC panel, 12 000SEK

Cell stocks 48x300k cells, small QC panel, 17 000SEK

Cell stocks 20x300k cells, large QC panel, 18 000SEK

Cell stocks 48x300k cells, large QC panel, 23 000SEK

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

small (1 plate), 1 800 SEK, delivers around 90ul of 10⁷-10⁸ TUI/ml*
medium (3 plates), 4 000 SEK, delivers around 300ul of 10⁷-10⁸ TUI/ml*
large (10 plates) 10 000 SEK, delivers around 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.

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

LV order form 2023.pdf

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): 11 000SEK
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.

AAV order form.pptx

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