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 vials.
Only 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.
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 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
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.
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.