Providing iPSC Generation and Characterization Services to Researchers

Initially described in the pioneering work of Yamanaka and colleagues, the ability to "reprogram" differentiated somatic cells into a pluripotent embryonic stem cell-like state by retroviral mediated expression of four specific transcription factors has revolutionized our ability to develop new models to study humans disease and represents a significant step toward patient-specific cell replacement therapies.

What can I do with iPSCs?

In addition to solving the ethical concerns related to the use of human blastocyst-derived embryonic stem cells, because it is possible to reprogram somatic cells derived from any individual, iPSC technologies provide a unprecedented tool for studying normal and pathological human development, and for developing novel models of human disease, including analysis of the effects of genetic variation on disease pathophysiology. In addition, iPSC-derived cells/tissues can be used for the discovery and validation of new therapeutic drug targets for treating human disease, and for identifying human-specific toxicities of drug candidates. Finally, the potential clinical application of cells/tissues derived from human iPSCs is also being evaluated in a number of clinical trials.

iPSC technologies we have used

• We have successfully generated iPSCs from numerous cell types.
• We currently use a non-integrating recombinant Sendai virus to generate high quality iPSCs from both fibroblasts and peripheral blood mononuclear cells with high efficiency.
• We have also used a non-integrating, episomal plasmid-based method and an excisable, polycistronic lentiviral vector to generate high quality, integration-free iPSCs.
• Using these methods, we have successfully generated high-quality iPSCs from hundreds of individuals with a broad range of diseases. Examples include:
• Pulmonary alveolar proteinosis
• Steatohepatitis
• Gaucher disease
• We have successfully generated iPSCs from peripheral blood mononuclear cells, fibroblasts, keratinocytes, lymphoblastoid cell lines (LCLs), CD34 bone marrow and cord blood, nasal and lung epithelial cells, and kidney epithelial cells derived from urine.

iPSC services available 

We provide the following iPSC services:

• Fee-for-service generation of patient-specific iPSC.
• Training in iPSC culture and generation (available only to Cincinnati Children's and University of Cincinnati investigators).
• Cryopreservation of patient-derived peripheral blood mononuclear cells for future iPSC generation.
• iPSC characterization and quality control assays.
• CRISPR/Cas-mediated gene editing of iPSCs.
• iPSC transduction with lentiviral/retroviral vectors (e.g. introduction of a gRNA expression cassette) and clone generation.
• Access to validated, robustly quality-controlled iPSCs derived from healthy and diseased individuals.

Streamlined patient consenting process

• Cincinnati Children's IRB-approved protocol covering any Cincinnati Children's patient for iPSC generation.
• Option for non-Cincinnati Children's patients to be consented.
• Available to both internal and external investigators.
• Remote consenting requires teleconference that must include the physician and participant or legal guardian if minor.
• Consented participants agree to all of the following: blood, saliva, skin biopsy (if surgical procedure is occurring) and review of medical records from Cincinnati Children's and outside hospitals.