Training Opportunities for Internal and External Scientists

The Pluripotent Stem Cell Facility offers a variety of hands-on training courses for internal and external scientists.

Basic Training

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. Training in the basic culture of hESCs/iPSCs is advised for individuals wishing to perform experiments in our facility and/or for those planning on ultimately transferring the technology to their own laboratories.

Our training course focuses on feeder-free methods for hPSC culture using the mTeSR1/Matrigel culture system, and covers all aspects of routine hPSC culture, including:

• Media/substrate preparation
• Routine hPSC maintenance
• Passaging pluripotent hPSCs
• hPSC cryopreservation and recovery
• The importance and methods for hPSC authentication and quality control

Additional Details

• The course is open to internal and external investigators.
• We can accommodate up to 3 people per lab for each training session.
• The cost of this training course includes ongoing access to online training videos, hPSCs, media, reagents, and a manual containing detailed protocols, as well as follow-up support from our technical staff.

Advanced Training

Generation of Human iPSCs

We offer a hands-on training course in the generation of human iPSCs. 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 (we currently use non-integrating sendai virus expression for reprogramming PBMCs and fibroblasts)
• Passaging transfected/transduced cells to feeders and culture (after ~1 week)
• Identification and excision of putative iPSC colonies (after 3-4 weeks)
• Expansion and cryopreservation of robust iPSC lines (4-6 weeks)

Additional Details

• The course is only open to Cincinnati Children's and University of Cincinnati investigators.
• The basic training in "Techniques for Human Pluripotent Stem Cell Culture" is a prerequisite for this course.
• Training in iPSC generation takes 3-4 months to complete but only involves attendance at our facility during a few 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. We can supply "normal" human fibroblasts or PBMCs if preferred.
• Cost includes media, all reagents and generation of up to 6 discrete clones from a single donor (6 cryovials per line). Cost does NOT include authentication and characterization of generated iPSC lines.

Generation of Human 3-dimensional Gastrointestinal Organoids

We also offer a hands-on training course in the generation of human intestinal organoids (HIOs), human antral gastric organoids (aHGOs), or human colonic organoids (HCOs) from hPSCs. Hands-on training covers all aspects of gastrointestinal organoid generation, including:

• Plating hPSCs
• Generation of definitive endoderm
• Endoderm patterning and formation and cryopreservation of posterior-foregut endoderm (PFE) or mid-hindgut endoderm (MHE)
• Thawing and aggregation of PFE or MHE
• Plating of PFE or MHE aggregates
• Generation of aHGOs, HCOs or HIOs
• Importance and methods for authentication and quality control

Additional Details

• The basic training in "Techniques for Human Pluripotent Stem Cell Culture" is a prerequisite for this course.
• Cincinnati Children's and University of Cincinnati trainees: HIO/HCO/aHGO training takes 7-8 weeks to complete but only involves attendance at our facility during a few key steps of the process.
• External trainees: We offer a 3-day condensed version of the training course covering all major steps in organoid generation for external trainees.
• Training costs include cells, media, all reagents and generation of 12 wells of organoids per trainee.

For more information about our courses, contact us.