iPS Core services

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.

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. 

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

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.