iPS Core services

Our services are tailor made after the requests from the customer. We can provide a full service chain from processing of dermal biopsies to fully differentiated neurons.

iPS core
Logo/iPS Core Facility/Karolinska Institutet

The iPS core facility provides reprogramming services to supply scientists with human induced pluripotent stem (iPS) cells, Neuroepithelial Stem (NES) cells and differentiated neurons. Additionally, we offer other services, such as cellbanking, training and consulting.

In collaboration with other core facilities, we have pipelines for establishment of genome-edited iPS cells using CRISPR-Cas9 technology. 

Fibroblast Derivation

Establishment of fibroblast cells from skin biopsy

This service takes about 1.5 months to complete and it includes:

  • Establishing fibroblast cells from a 2-4 mm skin biopsy
  • Feeding and splitting to deliver one T75 flask or three frozen vialsOnly freshly collected biopsies will be accepted. In the unlikely event that we fail to isolate fibroblasts from properly collected skin biopsies due to errors by the facility, there will be no charge for this attempt. We will ask the investigator to provide a new biopsy to complete the service.

iPS Reprogramming

mRNA-mediated reprogramming

Reprogramming human fibroblast cells using synthetic mRNA 

This service takes about 2-3 months and it includes:

  • Mycoplasma testing, expansion, cryopreservation and set-up of fibroblast cells for reprogramming.
  • Reprogramming using virus-free, synthetic mRNA.
  • Re-plating of transduced cells onto one 6cm plate with human recombinant laminin coating, daily feeding.
  • Manual picking of minimum 12 good looking colonies, 2-3 weeks after initiation of reprogramming.
  • Expansion of top four colonies, feeding daily.
  • Basic characterisation of top two iPS cell clones, including cell authentication, morphology by bright-field imaging and expression of pluripotency markers by FACS.
  • Delivery of 3-5 frozen vials of iPS cells of each clone.

 

iPS OCT4.
OCT4 expressing human iPS cells

iPS adaptation to LN521 and E8 medium condition

Transfer and adaptation of iPS cell lines to LN521 and E8 culture conditions

Laminin521 and E8 are defined culture conditions for pluripotent stem cells and we can adapt your iPS cell lines to these conditions.

This service takes 1-2 months and it includes:

  • Thawing the iPS cell lines in original culture conditions
  • Mycoplasma testing
  • Adaptation to LN521 and E8
  • Expansion, passaging and morphology confirmation
  • Delivery of 3-5 vials

Neural Induction

Neuroepithelial stem (NES) cells

NES cell lines can be maintained for >100 passages and make up a stable source material for differentiation to neurons and glia.

This service takes about 1-2 months and includes:

  • Neural induction of iPS cells in monolayer by dual-SMAD inhibition protocol.
  • Establishment of one stable neural precursor cells/neuroepithelial stem (NES) cell line from one iPS cell clone.
  • Expansion of NES cell lines in EGF/FGF2 for 6 passages.
  • Basic characterisation of NES cells, including cell authentication, morphology by bright-field imaging and expression of neural stem cell markers by FACS.
  • Delivery of 3-5 frozen vials of NES cells.
iPScore.
Human NES-derived Neurons-D#65

Neural Differentiation

Undirected differentiation of NES cells

By removing EGF/FGF2 from the culture, NES cells differentiate into 80-90% neurons and 10-20% glia after differentiation for 2 months or more.

This service takes up to 3 months (depends on customer demands) and it includes:

  • Thawing and expansion of NES cells.
  • Setting up for differentiation.
  • Daily feeding and adding of coating over the duration of the differentiation.
  • Basic characterisation of the differentiation, including morphology by bright-field imaging.
  • Sampling up on request.

Genome-edited iPS cell lines

Establishment of isogenic iPS cell lines using CRISPR-Cas9 systems

In collaboration with the Karolinska Genome Engineering (KGE) core facility at Karolinska Institutet, we establish iPS cell lines with edited genomic region of your interest.

This service takes approximately three-four months and it includes:

  • Consulting meeting together with KGE for feasibility
  • Coordination and experimental planning
  • Thawing and expansion of iPS cells
  • Preparation and conduct nucleofection
  • Recovering treated iPS cells and expansion
  • Confirming editing efficiency in bulk (ddPCR conducted by KGE facility)
  • Single-cell expansion based on editing efficiency
  • Delivering of single-cell clones for sequencing (performed by KGE facility)
  • Reporting the data and delivering edited iPS cell lines

Available Cell Lines

iPS cell lines and NES cell lines from non-traceable and non-diagnosed healthy control individuals.

We have iPS and NES cell lines available for requests and distribution. For details, see below. 

iPScore.
Cell storage in liquid nitrogen

Cell Banking

Safe storage of cell vials.

This service includes storage of fibroblast cells, iPS cells and NES cells in vapor phase liquid nitrogen tank for 1-10 years.

Karyotyping

Detection of chromosomal  abnormalities

G-banding, (Giemsa stain), traditional karyotyping method that can detect chromosomal abnormalities with a resolution down to 3Mb.

This service includes:

  • Expansion of cells
  • Preparation and fixation
  • Sending samples for G-banding
  • Reporting the data

Array-CGH (Comparative Genomic Hybridisation with Microarray), a technology that can detect unbalanced chromosomal abnormalities with a resolution down to 100kb.

This service includes:

  • Expansion of cells
  • Preparation gDNA
  • Sending samples to a core facility of your choice

EB formation

Pluripotency assay

Embryonic Body (EB) formation and germ layer analysis with RT-PCR. This shows if an iPS cell line has the potential to differentiate into the three germ layers.

This service will take approximately one month and it includes:

  • Expanding iPS cells
  • Setup EB formation
  • Harvesting EB s after two weeks
  • RNA preparation and RT-PCR with markers for all three germ layers
  • Reporting results as gel pictures
training
Hands-on Training at iPS core facility

Cell Expansion (freezing-thawing)

Providing vials of established fibroblasts, iPS and NES cells

This service includes thawing, expansion and freezing of cells to deliver the desired quantity of cryo-vials.

Sample preparation for DNA/RNA

Isolating DNA/RNA from fibroblasts, iPS cells, NES cells and neurons

Isolated DNA or RNA can be used for example for bulk and single-cell sequencing, PCRs and arrays. This service includes:

  • Thawing and expansion of cells
  • Preparation of DNA and RNA
  • Delivery to customer or a chosen core facility at SciLifeLab or Karolinska Institutet

Training

iPS Culture

Human iPS cell culture training

The training is tailored for each customer, in terms of duration, content and dates. You will be guided by an experienced lab technician on the basic culturing techniques such as thawing, passaging and freezing iPS cells under feeder-free condition.

NES Culture

Human NES cell culture and differentiation training

The training is tailored for each customer, in terms of duration, content and dates. You will be guided by an experienced lab technician on culturing, freezing and thawing proliferating NES cells as well as differentiation of them to neurons and glia.

Consulting

Consulting on project planning and design, financial planning, ethical considerations and result interpretation.

Price list for the services above

The listed prices are excluded INDI. INDI of 23% will be added to all prices.

This price list is valid until December 31 2021.

Price List 2021 - Fibroblasts

Price List 2021 - Fibroblasts
SEK Unit
Fibroblast derivation 9,600.00 per line for 3-5 vials
Expansion/freezing-thawing 3,000.00 for 3 vials
Cell Storage 500.00 per vial (yearly fee)
Mycoplasma test 500.00 per sample
Karyotyping 5,500.00 per sample

Price List 2021 - iPS 

Price List 2021 - iPS
SEK Unit
Request for available iPS cell lines 5,500.00 for 3 vials
iPS reprogramming 59,000.00 per line for 3-5 vials
Adaptation to LN521 and E8 medium condition 7,200.00 per line
Expansion/freezing-thawing 5,500.00 for 3 vials
Cell Storage 500.00 per vial (yearly fee)
Karyotyping 5,500.00 per sample
FACS (with analyzer-only) - Included in reprogramming service
CRISPR-edited iPS lines 59,000.00 for 3-5 vials
Sample prep for DNA/RNA 6,000.00 per sample
EB formation 7,000.00 per sample

Price List 2021 - NES

Price List 2021 - NES
SEK Unit
Request for available NES cell lines 5,500.00 for 3 vials
Neural Induction 59,000.00 per line for 3-5 vials
Expansion/freezing-thawing 5,500.00 for 3 vials
Cell Storage 500.00 per vial (yearly fee)
FACS (with analyzer-only) - Included in neural induction service
Sample prep for DNA/RNA 6,000.00 per sample
Neural Differentiation 8,400.00 per (full) 6-well-plate

Price List 2021 - Training

Price List 2021 - Training
SEK Unit
Consulting 1,500.00 per hour
Training 1,000.00 per hour

Available iPS Cell Lines

Available iPS cell lines
Cell Line Gender Age Reprogramming Method Reference (PMID)
CTRL5 Female Child Sendai Virus 25043502
CTRL7 Male Middle Age Sendai Virus 27789397
CTRL9 Male Child Sendai Virus/mRNA 28395796
CTRL10 Male Child Sendai Virus/mRNA 28395796
KICR001-A Male Old mRNA
CTRL14 Female Child mRNA

Available NES Cell Lines

Available NES cell lines
Cell Line Gender Age Comment Reference (PMID)
CTRL1 Female Old 28924182
CTRL3 Male Child 28924182
CTRL5 Female Child
CTRL7 Male Middle Age 31302032
CTRL9 Male Child 29456185
CTRL10 Male Child 31302032
KICR001-A Male Old
AF22 Female Middle Age 22272239
Sai2 Captured from the developing brain 23884946