Establishment of Marmoset iPS-derived Cranial Neural Crest Cells

狨猴 iPS 来源的颅神经嵴细胞的建立

基本信息

批准号：
9085180
负责人：
Steven Farnsworth
金额：
$ 4.36万
依托单位：
UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-08-01 至 2016-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9085180
关键词：
Address Adult Afferent Neurons Animal Model Autologous Biochemical Markers Biological Models Biomedical Research Bone Regeneration Callithrix Callithrix jacchus jacchus Cartilage Cell Differentiation process Cell Lineage Cell Maintenance Cell Therapy Cell Transplantation Cell model Cells Cephalic Clinical Complex Cranial Nerves Cues Dental Dental Pulp Dentin Dentistry Dentists Development Electrophysiology (science)Embryo Embryonic Development Epithelial Epithelium Event Face Flow Cytometry Future Gene Expression Generations Genes Goals Human Human Development In Vitro Investigation Laboratories Mandible Maxilla Meckel&apos s cartilage Mesenchymal Mesenchymal Differentiation Mesenchyme Modeling Molecular Molecular Profiling Multipotent Stem Cells Natural regeneration Neural Crest Neural Crest Cell Neurons Odontoblasts Osteoblasts Patients Peripheral Peripheral Nerves Peripheral Nervous System Phenotype Play Pluripotent Stem Cells Population Pregnancy Primates Research Research Proposals Role Safety Schwann Cells Scientist Signal Pathway Signal Transduction Somatic Cell Sorting - Cell Movement Source Specific qualifier value Stem cells Structure Structure of trigeminal ganglion Testing Tissue Transplantation Tooth structure Training Translational Research bone career cell motility cell type clinical application extracellular face bone structure immunocytochemistry in vivo induced pluripotent stem cell innovation nonhuman primate osteogenic regenerative relating to nervous system stem stem cell population therapy development tissue regeneration

项目摘要

DESCRIPTION (provided by applicant): 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.

描述（由申请人提供）：成体细胞可以被重新编程为胚胎样诱导多能干（iPS）细胞的突破性发现为未来创新的患者特异性细胞和组织再生疗法带来了兴奋。 iPS 细胞在适当的条件下可以发育成任何所需的细胞类型。对于再生牙科目标，从 iPS 细胞衍生颅神经嵴 (CNC) 细胞是未来开发 iPS 衍生颅面部结构再生疗法的重要一步。为了评估 iPS 衍生细胞疗法的安全性和有效性，非常需要开发非人类灵长类动物模型系统。普通狨猴 (Callithrix jacchus) 是一种广泛用于生物医学研究的非人类灵长类动物，最近从狨猴中产生了 iPS 细胞。拟议的研究将探讨狨猴诱导多能干（iPS）细胞在体外发育颅神经嵴细胞的潜力，并评估其体外下游分化潜力。 CNC 细胞是多能干细胞，在脊椎动物发育过程中产生颅雪旺细胞、周围神经元、面骨、软骨和牙齿发生所需的牙齿间质。 iPS 衍生的 CNC 细胞及其下游谱系特异性后代细胞是牙齿再生目标的理想细胞，例如骨骼、牙本质、牙髓、全牙和周围神经的再生。狨猴 iPS 衍生的 CNC 细胞的开发将加速人类 iPS 衍生疗法的开发，通过非人类灵长类细胞治疗模型进行颅面部结构再生。我们的总体假设是狨猴 iPS 细胞将产生具有神经和间充质细胞谱系分化潜力的颅神经嵴干细胞。该假设将通过以下具体目标进行检验：1）建立狨猴 iPS 衍生的 CNC 细胞，2）确定狨猴 iPS 衍生的 CNC 细胞在体外的神经分化潜力，3）确定狨猴的间充质分化潜力体外 iPS 衍生的 CNC 细胞。这些研究将提供 iPS 重编程、干细胞维持和分化、定量基因表达、免疫细胞化学、流式细胞术、FACS 分选、动物建模和神经元电生理学方面的培训。该提议具有创新性，因为尚未使用非人灵长类多能干细胞证明 CNC 分化。这项研究意义重大，因为 CNC 细胞是定义适当的早期颅面部发育的关键群体。这些研究将通过分离 CNC 细胞和富集 CNC 衍生细胞类型（如雪旺细胞前体细胞、成骨细胞和成牙本质细胞）来影响用于颅面结构再生的 iPS 衍生细胞疗法的开发。未来狨猴中的自体细胞疗法将通过评估体内细胞疗法的长期安全性和有效性，影响 iPS 衍生疗法的发展。该研究计划符合申请人成为一名研究再生牙科转化研究的牙医科学家的职业目标。