Pluripotent Stem Cell Facility

Pluripotent Stem Cell Facility

Induced Pluripotent Stem Cells (iPSCs)

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 human disease and represents a significant step toward patient-specific replacement therapies.

What can I do with iPSCs?

In addition to solving ethical concerns related to the use of blastocyst-derived embryonic stem cells, the use of iPSCs for the generation of therapeutic cells for cell replacement therapy may avoid the requirement for post-transplant immune suppression because iPSCs can be generated directly from the transplant recipient and will therefore be genetically identical to the patient. Additionally, because it is possible to reprogram somatic cells derived from diseased individuals, iPSC technology provides an important new platform for the development of new models of human disease. Thus, upon appropriate differentiation, these cells can then be used to study normal and pathologic human tissue development in vitro, enabling new insights into disease pathology as well as the development of novel therapeutic agents and patient-specific cell replacement therapies.

iPSC Technologies Used in the PSCF

• We have successfully generated iPSCs using multiple methods of reprogramming factor expression in numerous cell types.
• Currently, the PSCF uses a non-integrating recombinant Sendai virus to generate high quality iPSCs from both fibroblasts and non-mobilized peripheral blood 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 also successfully generated high-quality iPSCs from disease-specific fibroblasts and peripheral blood mononuclear cells.
• 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 in the PSCF

The PSCF is committed to facilitating access to this exciting technology for Cincinnati Children's / UC researchers as well as external investigators.
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 / UC 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
• Access to existing quality-controlled iPSCs derived from healthy individuals

Streamlined patient consenting process

• Cincinnati Children's Hospital Medical Center IRB-approved protocol covering any Cincinnati Children's Hospital Medical Center patient for iPSC generation
• Option for parents and other non-Cincinnati Children's Hospital Medical Center patients to be included
• 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 Hospital Medical Center and outside hospitals.