Reference Transcriptome for Deriving Marmoset Induced Pluripotent Stem Cells
Limited access to primary tissue from various nonhuman primate (NHP) species represents a significant unmet need that hampers progress in understanding unique cellular diversity and gene regulation of specific tissues and organs in stem cell translational research. Most comparative biology studies have been limited to using postmortem tissue usually frozen specimens with limited utility for research. The generation of induced pluripotent stem cell (iPSC) lines from somatic cells, such as adult skin or blood cells, offers an alternative to invasive and ethically controversial interventions for acquiring tissue. Pluripotent iPSCs have virtually an unlimited capacity to proliferate and differentiate into all cell types of the body. We are generating high-quality validated NHP iPSC lines to offer to scientific community and facilitate their research programs. We use the non-integrative episomal vector system to generate iPSCs from NHP skin biopsies. In this chapter we describe the validation of NHP iPSC lines by confirming pluripotency and their propensity to differentiate into all three germ layers ectoderm, mesoderm, and endoderm according to established standards and measurable limits for a set of marker genes incorporated into a scorecard.
Key wordsMarmoset iPSCs Pluripotency Scorecard iPSC characterization Nonhuman primate iPSCs Regenerative medicine
The authors thank members of the Daadi laboratory for helpful support and suggestions. This work was supported by the Worth Family Fund, The Perry & Ruby Stevens Charitable Foundation, The Robert J. Kleberg, Jr., and Helen C. Kleberg Foundation, the NIH primate center base grant (Office of Research Infrastructure Programs/OD P51 OD011133), and the National Center for Advancing Translational Sciences, National Institutes of Health, through Grant UL1 TR001120.
Disclosures: Dr. Marcel M. Daadi is founder of the biotech company NeoNeuron.