Services

We reprogram skin fibroblasts with the following vector systems:

1. Episomal plasmids (Okita et al. 2011); occasionally integrating into host genome
2. Sendai virus (SeV, Cytotune kit 2.0, Thermo Fisher); integration-free
3. Self-replicating RNA (ReproRNA kit, Stem Cell Technologies); integration-free

We need 5x 10⁵  fibroblasts to begin with. They must be highly proliferative and should have the lowest passage number possible.

We reprogram erythroblasts which we enrich from peripheral blood mononuclear cells (PBMCs). We can either use cryopreserved PBMCs (at least 2x10^6 live cells) or we can isolate PBMCs from fresh blood (at least 10 ml, not older than 24h).

We reprogram erythroblasts with the following vector systems:

1. Episomal plasmids (Okita et al. 2011); occasionally integrating into host genome
2. Sendai virus (SeV, Cytotune kit 2.0, Thermo Fisher); integration-free

We can isolate urine-derived cells from >150 ml urine; urine must be processed on the day of donation.

We reprogram urine-derived cells with the following vectors systems:

1. Episomal plasmids (Okita et al. 2011); occasionally integrating into host genome
2. Sendai virus (SeV, Cytotune kit 2.0, Thermo Fisher); integration-free
3. Self-replicating RNA (ReproRNA kit, Stem Cell Technologies); integration-free.

We have several ‘control’ hiPSCs available which have been extensively characterized:

• from male and female donors
• with an updated (2025) informed consent allowing various research applications
• generated from different primary cells and with different reprogramming vectors
• with additional characterization (e.g. HLA typing, X-chromosome status, WGS)
• used by multiple researchers in various research projects (publication list available)
• registered in hPSCreg  

We can genetically modify your hiPSC line of choice with CRISPR/Cas9 for the generation of:

• Isogenic hiPSCs by either converting the disease-causing mutation into the WT sequence or by introducing a mutation into a gene of interest in a ‘control’ hiPSC line.
• Knockout hiPSCs by partial deletion of a crucial exon
• Reporter hiPSCs by tagging your gene of interest with a fluorescent protein

After electroporation with Cas9 protein we clone hiPSCs and screen resulting hiPSC colonies for successful editing with PCR/ restriction enzyme analysis followed by Sanger sequencing. Up to 6 clones are expanded for cryopreservation. You can choose the number of successfully edited clones for characterization (see Service 6).   

Based on the requirements of many peer-reviewed journals for newly generated hiPSCs or genetically modified hiPSCs we offer the following standard QC service:

• Morphology: brightfield images at low magnification to assess typical features of undifferentiated hiPSCs (e.g. defined colony edges; homogenous population of undifferentiated cells)
• Pluripotency status: Quantitative analysis of markers of undifferentiated state (OCT 3/4, Nanog, SSEA-4, Tra-1-60) by flow cytometry
• Functional pluripotency: Short-term in vitro differentiation into Ecto-, Meso-, and Endoderm followed by qualitative immunofluorescent staining analysis (3 markers per germ layer)
• Genetic integrity: CNV analysis of the most common abnormalities by ddPCR (Stem Genomics iCS digital service)
• Cell identity: Short tandem repeat (STR) analysis is used to confirm identity of hiPSCs (and primary cells, if available; service provided by LUMC dept HG)  

The STRAIGHT-IN platform is designed for facile genomic integration of DNA payloads (up to 50kb) into hiPSCs that contain a pre-inserted landing pad in a safe harbor.

Two control hiPSC lines (male and female donor) have been equipped with STRAIGHT-IN landing pads in safe harbor loci AAVS1 or CLYBL. A hiPSC line with two landing pads which can be targeted individually is also available. Additional STRAIGHT-IN versions are currently being developed; contact us for details.

1. Annual training course in collaboration with Stem Cell Technologies: 2.5-day course on maintenance of undifferentiated hiPSCs and basic principles of differentiation. Hands-on training includes passaging, freezing and thawing of undifferentiated hiPSCs as well as examples for 2D and 3D differentiation. Extensive theoretical background is provided during lectures. For researchers with little experience or for refreshing your memories. Contact us for dates!
2. Training courses on more specialized topics such as vascular cells and microfluidics, gene editing etc. Organized from time to time. Contact us for more information.
3. Customized trainings upon request: Would you like to get specific support e.g. for differentiation of hiPSCs into your cell type of interest based on a publication by other researchers? Contact us and let’s discuss how we can help you!