Services Provided by the Core
The Bioengineering Core will generate custom-designed iPSC from primary mammalian cells (e.g. human, mouse, dog). A major focus of our Core is the generation of iPSC from human patients with inherited diseases. The Core will isolate, amplify and freeze primary fibroblasts from skin biopsies to be used for the generation of iPSC. Other tissue and cell sources can be used as well (inquire). Up to 5 cell clones per iPSC conversion will be provided to the investigators. Our standard method to reprogram primary cells into iPSC utilizes Sendai Virus Vectors. These vectors do not integrate into the iPSC genome and thus do not result in a genetic footprint. Alternatively, we are offering a RNA-based reprograming system. Our baseline iPSC characterization package includes mycoplasma testing, QRT-PCR assessment of stem cell marker expression and karyotyping (inquire about details). 3 vials of frozen iPSC per clone will be provided to the investigator. Please note that additional services are offered to characterize iPSC, including in vitro differentiation assays (to assess the ability to differentiate into cells representing all three germ layers; see main menu). It might take up to 3 months to complete an iPSC project. The Core will release iPSC that meet basic quality control criteria, such as expression of a set of pluripotency markers as judged by qRT-PCR (inquire for details).
Responsibilities of the Investigator
- Discuss experimental design with Bioengineering Core Director.
- Provide primary cells for iPSC reprograming. The Core can isolate fibroblasts from skin biopsies. Other sources of cells are acceptable (please inquire).
- Provide proof that cells were obtained with all required regulatory approvals.
- Provide information (if available) as to whether human samples carry infectious diseases. The facility will not accept samples carrying pathogens which require biosafety level 2 handling.
- Sign an agreement regarding acceptable use of Core-generated iPSC.
Additional Characterization of iPSC (inquire about details)
- QRT-PCR, RT-PCR or ICC assessment of pluripotency marker expression
- Karyotyping analysis
- In vitro embryoid body assay to assess the potential of iPSC to spontaneously differentiate into cells representing all three germ layers.
- iPSC clone expansion and freezing of up to 10 vials.
- Mycoplasma testing