Stem Cell Facility

We generate feeder-free induced pluripotent stem cell (iPSC) lines from human blood, skin fibroblasts, and potentially other cell-types.

Current IPSC Approach

• CytoTune-iPS 2.0 Sendai Reprogramming Kit (ThermoFisher Scientific).

Advantages

Our current iPSC approach has several advantages, including:

• it efficiently reprograms blood and other cell types,
• it has scalable cell line generation with minimal hands-on time,
• and it has a non-integrating cytoplasmic RNA vector that rapidly clears.

iPSC Characterization Services

• Immunostaining of pluripotent markers.
  • Surface markers: TRA-1-60 and SSEA4.
  • Nuclear markers: Nanog, Oct4, Sox2.
• qPCR analysis of pluripotent markers.
  • Core pluripotent markers upregulated: Nanog, Pou5f1, Sox2, Tfcp2L1.
  • Blood markers silenced: Cd3d, Cd4, Cd8a, Ccr7.
  • Reference markers: Rplp0, Sdha.
• Mycoplasma and bacteria PCR-based testing (part of a continuous testing plan when generating iPSC lines).
• Validation of pluripotency: Stemdiff Trilineage Differentiation Kit.
• Flow Cytometry of pluripotent markers
  • Surface markers: TRA-1-60 and SSEA4.
  • Nuclear markers: Nanog, Oct4, Sox2.

iPSC Differentation Services

Methods for differentating IPSCs into a variety of lineages are improving and developing at a rapid pace. Currently we use established protocols that efficiently convert iPSCs into cardiomyocytes/cardiovascular cells. We are exploring options for generating neural lineages. Other cell-type specific protocols may follow. We welcome any inquiries into available resources and will provide basic advice on how to prep iPSCs for differentiation into desired lineages.