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Functional characterization of adult taste stem cells

成体味觉干细胞的功能表征

基本信息

批准号：
9086574
负责人：
PeiHua Jiang
金额：
$ 0.59万
依托单位：
MONELL CHEMICAL SENSES CENTER
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-04-01 至 2020-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9086574
关键词：
3-Dimensional Address Adult Affect Anterior Area Biological Assay Boxing Cancer Patient Cell Culture Techniques Cell Lineage Cells Circumvallate Papilla Cues Culture Media Development Diabetes Mellitus Diet Disease Erinaceidae Fluorescence-Activated Cell Sorting Foliate Papilla Genetic Genetically Engineered Mouse Goals Growth Head and neck structure Health Homologous Gene Hypertension Immunohistochemistry Impairment In Vitro Individual Intestines Lead Leucine-Rich Repeat Mus Natural regeneration Nature Obesity Organoids Patients Population Process Proliferating Protocols documentation Radiation therapy Regimen Reporter Research Reverse Transcriptase Polymerase Chain Reaction Route Signal Transduction Sodium Chloride Sorting - Cell Movement Stem cells Structure System Taste Bud Cell Taste Buds Taste Perception Testing Time Tissues Tongue Transgenes Transgenic Organisms Villus base cell type chemotherapy enhanced green fluorescent protein human CCXCR1 receptor in vivo matrigel multipotent cell novel regenerative release of sequestered calcium ion into cytoplasm research study screening self-renewal stem stemness sugar tongue papilla

项目摘要

DESCRIPTION (provided by applicant): This proposed research will examine how adult taste stem cells generate mature taste cells, and will develop a culture system to grow, expand, and differentiate taste stem cells to study taste regeneration processes in vitro. The nature of taste stem cells is unknown although those in the posterior tongue express Lgr5; those in both the anterior and posterior express Lgr6.To expand our preliminary understanding of these cells, we propose to use genetic lineage tracing strategies and to establish a novel cell culture approach to further characterize Lgr5+ and Lgr6+ taste cells and determine their regenerative capacity both in vivo and in vitro. In this project, we will investigate two overlapping areas: In Aim 1, we will determine whether Lgr5+ and Lgr6+ cells are taste stem/progenitor cells in vivo. We will characterize the nature of Lgr5+ and Lgr6+ cells in detail using existing genetically engineered mice and various marker induction regimens. Subaims will determine whether (Aim 1.1) the strong Lgr5+ cells are taste stem cells for posterior tongue, (Aim 1.2) Lgr5+ cells give rise to Shh+ cells en route to mature taste bud cells, (Aim 1.3) individual taste buds are monoclonal (derived from a single progenitor cell) or oligoclonal (derived from a few progenitor cells) and Lgr5+ cells are multipotent (differentiate into different cell types) or unipotent (develop into ony one cell type), and (Aim 1.4) Lgr6+ cells represent adult taste stem/progenitor cells in anterior and posterior tongue. In Aim 2, we will determine the regenerative capacity of Lgr5+ and Lgr6+ cells in vitro. Single Lgr5+ intestinal stem cells are able to build crypt-villus structures in vito. We will isolate Lgr5+ and Lgr6+ stem cells from tongue by FACS sorting and culture them in defined medium in ultra-low attachment plates to examine their growth, expansion, and long-term passages (Aim 2.1), as well as the relatedness of Lgr5+ and Lgr6+ cells. Using a Matrigel-based 3-D culture system, we will determine the regenerative capacity of Lgr5+ and Lgr6+ cells, whether single Lgr5+ and Lgr6+ cells are multipotent or unipotent, and which culture components affect growth, expansion, and differentiation (Aim 2.2). These areas of study are significant first steps toward a regenerative approach for treating patients who suffer from taste impairment, such as cancer patients receiving head and neck radiation therapy or chemotherapy, and may lead to the development of a culture system for screening novel taste modulators.