The groundbreaking discovery that adult cells can be reprogrammed into embryonic-like induced pluripotent stem (iPS) cells has developed excitement for future innovative patient-specific cell and tissue regeneration therapies. iPS cells can develop into any desired cell type under appropriate conditions. For regenerative dentistry goals, deriving cranial neural crest (CNC) cells from iPS cells is an important step for future development of iPS-derived cranial-facial structure regeneration therapies. In order to assess the safety and efficacy of iPS-derived cell therapies, the development of a non-human primate model system is highly desirable. The common marmoset (Callithrix jacchus) is a non-human primate used widely in biomedical research and iPS cells were recently generated from the marmoset. The proposed studies will address the potential of marmoset induced pluripotent stem (iPS) cells for in vitro development of cranial neural crest cells and assess their downstream differentiation potential in vitro. CNC cells are multipotent stem cells that during vertebrate development give rise to cranial Schwann cells, peripheral neurons, facial bone, cartilage, and the dental mesenchyme required in tooth genesis. iPS-derived CNC cells and their downstream lineage-specific progeny cells make ideal cells for dental regenerative goals such as the regeneration of bone, dentin, dental pulp, whole teeth, and peripheral nerves. The development of marmoset iPS-derived CNC cells will accelerate the development of human iPS-derived therapies for cranial-facial structure regeneration through non-human primate cell therapy modeling. Our overall hypothesis that marmoset iPS cells will generate cranial neural crest stem cells with differentiation potential in both neural and mesenchymal cell lineages. This hypothesis will be tested by the following specific aims: 1) to establish marmoset iPS-derived CNC cells, 2) to determine the neural differentiation potential of marmoset iPS-derived CNC cells in vitro and 3) To determine the mesenchymal differentiation potential of marmoset iPS-derived CNC cells in vitro. These studies will provide training in iPS reprogramming, stem cell maintenance and differentiation, quantitative gene expression, immunocytochemistry, flow cytometry, FACS sorting, animal modeling, and neuron electrophysiology. This proposal is innovative because CNC differentiation has not been demonstrated using non-human primate pluripotent stem cells. The proposed research is significant because CNC cells are a key population that defines proper early cranial-facial development. These studies will impact the development of iPS-derived cell therapies for cranial-facial structure regeneration through the isolation of CNC cells and enrichment of CNC-derived cell types such as Schwann cell precursors, osteoblasts, and odontoblasts. Future autologous cell therapy in the marmoset will impact the development of iPS-derived therapies by assessing the long-term safety and efficacy of cell therapy in vivo. This research proposal fits the applicant's career goal to become a dentist-scientist studying translational research in regenerative dentistry.

Public Health Relevance

Induced pluripotent stem (iPS) cells are a new and exciting source of cells useful to develop stem cell populations that will promote the regeneration of tissues and structures in the body through cell or tissue transplantation. Cranial neural crest (CNC) stem cells are an ideal population to develop from iPS cells due to their known critical role in contributing to the proper formation of cranial-facial cartilage, bone, teeth, and the cranial peripheral nervous system during vertebrate development. The planned studies will develop and characterize CNC stem cells from iPS cells of the common marmoset (Callithrix jacchus);a non-human primate model useful for developing and evaluating stem cell based therapies.

National Institute of Health (NIH)
National Institute of Dental & Craniofacial Research (NIDCR)
Individual Predoctoral NRSA for M.D./Ph.D. Fellowships (ADAMHA) (F30)
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NIDCR Special Grants Review Committee (DSR)
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Frieden, Leslie A
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University of Texas Health Science Center San Antonio
Schools of Medicine
San Antonio
United States
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Mishra, Anuja; Qiu, Zhifang; Farnsworth, Steven L et al. (2016) Induced Pluripotent Stem Cells from Nonhuman Primates. Methods Mol Biol 1357:183-93
Qiu, Zhifang; Mishra, Anuja; Li, Miao et al. (2015) Marmoset induced pluripotent stem cells: Robust neural differentiation following pretreatment with dimethyl sulfoxide. Stem Cell Res 15:141-50
Farnsworth, Steven L; Qiu, Zhifang; Mishra, Anuja et al. (2013) Directed neural differentiation of induced pluripotent stem cells from non-human primates. Exp Biol Med (Maywood) 238:276-84