The Pluripotent Stem Cell Facility offers a variety of training for internal and external scientists.
Training course in techniques for human pluripotent stem cell culture
The maintenance and manipulation of high-quality human pluripotent stem cells in culture is a relatively complex and labor-intensive process. The PSCF offers hands-on training in these techniques to CCHMC and UC scientists as well as external investigators.
Training in the basic culture of hESCs/iPSCs is advised for individuals wishing to perform experiments in the PSCF and/or for those planning on ultimately transferring the technology to their own laboratories.
The PSCF training course focuses on feeder-free methods for hPSC culture using the mTeSR1/matrigel culture system.
This course covers all aspects of routine hPSC culture, including:
- Routine hPSC maintenance
- Passaging pluripotent hPSCs
- hPSC cryopreservation and recovery
The course is open to both CCHMC/UC investigators and external investigators. We can accommodate up to a maximum of three people per lab for each training session.The cost of this training course includes cells, media, all reagents and training manual, as well as follow-up support from PSCF technical staff.
Please contact us for available dates, and internal investigators may also view pricing information.
Generation of human iPSCs
The PSCF now offers a hands-on training course in the generation of human iPSCs. This course is only available to CCHMC/UC investigators.
Hands-on training covers the whole process of iPSC generation from either fibroblasts or PBMCs, including:
Donor cell mycoplasma testing
- Media preparation and culture of donor-derived cells
- Expression of reprogramming factors (fibroblast: episomal plasmid or lentiviral expression available; PBMC: lentiviral expression available)
- Passaging transfected/transduced cells to feeders and culture in hESC media (after ~1 week)
- Identification and excision of putative iPSC colonies (after 3-4 weeks)
- Expansion and cryopreservation of robust iPSC lines (4-6 weeks)
The "Basic training course in techniques for pluripotent stem cell culture" is a prerequisite for this course
Training in iPSC generation takes 3-4 months to complete but involves attendance at the PSCF labs only at the key steps of the process.
To maximize the cost effectiveness of this training course, we recommend reprogramming disease-specific, donor-derived cells during training. Donor cells must be supplied by the trainee. The PSCF can supply "normal" human fibroblasts or PBMCs if preferred.
There is a small chance that an iPSC generation experiment will not produce stable iPSC lines (for non-technical reasons likely related to donor cell properties/genetics). Investigators will be charged a fee for each failed attempt. In this case, we suggest repeating the experiment using cells from a different donor if possible.
Cost includes media, all reagents and generation of up to 6 discrete lines from a single donor (6 cryovials per line). Cost does NOT include characterization of human iPSC lines.
Please contact us to discuss details and for available dates and times for training. Internal investigators may also view pricing information.
Generation of human intestinal organoids
The PSCF now offers a hands-on training course in the generation of human intestinal organoids (HIOs) from hPSCs. This course is available to both CCHMC/UC and external investigators.
Hands-on training covers all aspects of HIO generation, including:
- Plating hPSCs
- Generation of definitive endoderm
- Endoderm patterning and formation of hindgut spheroids
- Harvesting and plating of spheroids
- Generation and passaging of HIOs.
The "Basic training course in techniques for pluripotent stem cell culture" is a prerequisite for this course.
CCHMC/UC trainees: HIO training takes 7-8 weeks to complete but involves attendance at the PSCF labs only at the key steps of the process.
External trainees: a 3 day, condensed version of the HIO training course covering all major steps in HIO generation, is available for external trainees.
Training costs include cells, media, all reagents and generation of 12 wells of HIOs per trainee.
Please contact us for available dates and times for training. Internal investigators may also view pricing information.