Establishment of Marmoset iPS-derived Cranial Neural Crest Cells

狨猴 iPS 来源的颅神经嵴细胞的建立

• 批准号: 8256468
• 负责人: Steven Farnsworth
• 金额: $ 2.99万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8256468
• 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' 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

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. 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.

描述（由申请人提供）：开创性的发现，可以将成年细胞重新编程为类似胚胎的诱导多能茎（IPS）细胞已为未来的创新患者特异性细胞和组织再生疗法而产生了兴奋。在适当条件下，IPS细胞可以发展成任何所需的细胞类型。对于再生牙科目标，从IPS细胞中得出颅神经rest（CNC）细胞是IPS衍生的颅相结构再生疗法的未来开发的重要步骤。为了评估IPS衍生的细胞疗法的安全性和有效性，非常需要非人类灵长类动物模型系统的发展。常见的果酱（Callithrix jacchus）是一种非人类的灵长类动物，在生物医学研究中广泛使用，而IPS细胞最近是由果棒产生的。拟议的研究将介绍摩尔莫斯群诱导的多能干（IPS）细胞在体外发育颅神经rest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会产生颅施旺氏细胞，周围神经元，面部骨骼，软骨和牙齿生成所需的牙齿质质。 IPS来源的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，牙本质，牙髓，全牙和周围神经的再生。 Marmoset IPS衍生的CNC细胞的发展将通过非人类灵长类动物的细胞疗法建模加速颅IPS衍生疗法，用于颅面结构再生。我们的总体假设是Marmoset IPS细胞将在神经和间充质细胞谱系中产生具有分化潜力的颅神经rest干细胞。该假设将通过以下特定目的进行检验：1）建立Marmoset IPS衍生的CNC细胞，2）确定Marmoset IPS衍生的CNC细胞的神经分化潜力，并在体外和3）确定Marmoset的间隙分化潜力体外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，流式细胞仪，FACS分类，动物建模和神经元电生理学的培训。该建议具有创新性，因为使用非人类灵长类动物多能干细胞尚未证明CNC分化。拟议的研究很重要，因为CNC细胞是定义适当的早期颅面发育的关键人群。这些研究将通过分离CNC细胞并富集CNC衍生的细胞类型（例如Schwann细胞前体，成骨细胞和odontoblasts）来影响IPS来源的IPS衍生细胞疗法的发展，以通过分离CNC细胞和富集CNC衍生的细胞类型的发展。未来的摩尔马斯体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功效来影响IPS衍生的疗法的发展。这项研究建议符合申请人的职业目标，成为牙医科学家，研究再生牙科的翻译研究。 公共卫生相关性：诱导多能茎（IPS）细胞是一种有用的新的令人兴奋的细胞来源，可通过细胞或组织移植来促进体内组织和结构的再生。颅神经rest（CNC）干细胞是IPS细胞发展的理想人群，因为它们在脊椎动物发育过程中已知的关键作用在促进颅面软骨，骨骼，牙齿和颅神经周围神经系统的正确形成中的重要作用。计划的研究将开发并表征来自普通果棒IPS细胞（Callithrix jacchus）的CNC干细胞；一种非人类灵长类动物模型，可用于开发和评估基于干细胞的疗法。