Distinguishing adipose stromal vs. stem cells by serial transplantation

通过连续移植区分脂肪基质细胞和干细胞

基本信息

批准号：
8511619
负责人：
Bruce A. Bunnell
金额：
$ 23.92万
依托单位：
TULANE UNIVERSITY OF LOUISIANA
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：
2012-07-16 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8511619
关键词：
3-Dimensional Adherent Culture Adipocytes Adipose tissue Antigens Basic Science Biocompatible Blood Vessels Bone Marrow Bromodeoxyuridine CD29 Antigen CD34 gene Cell Culture Techniques Cell Fraction Cell Separation Cell Transplants Cell model Cells Characteristics Chondrogenesis Clinical Clinical Investigator Colony-Forming Units Assay Colony-forming units Commit Confocal Microscopy Diet Discipline Electron Microscopy Exhibits Fatty acid glycerol esters Flow Cytometry Gold Growth Factor Harvest Hematopoietic stem cells Histocytochemistry Human Immunodeficient Mouse Immunophenotyping Implant In Vitro Label Laboratories Laser Scanning Confocal Microscopy Life Literature MCAM gene Medical Modeling Monitor Mus Natural regeneration Obesity Organ Osteogenesis Outcome Pathway interactions Patients Physicians Population Process Regenerative Medicine Reporting Research Personnel Safety Semantics Silk Solid Sorting - Cell Movement Source Spleen Stem cell transplant Stem cells Surface Surface Antigens Testing Tissue Engineering Tissues Transgenes Transgenic Mice Transplantation adult stem cell base collagenase in vivo lipid biosynthesis mouse model progenitor public health relevance reconstitution regenerative research study scaffold self-renewal stem stem cell biology stem cell population stromal progenitor subcutaneous transcription factor

3-Dimensional Adherent Culture Adipocytes Adipose tissue Antigens Basic Science Biocompatible Blood Vessels Bone Marrow Bromodeoxyuridine CD29 Antigen CD34 gene Cell Culture Techniques Cell Fraction Cell Separation Cell Transplants Cell model Cells Characteristics Chondrogenesis Clinical Clinical Investigator Colony-Forming Units Assay Colony-forming units Commit Confocal Microscopy Diet Discipline Electron Microscopy Exhibits Fatty acid glycerol esters Flow Cytometry Gold Growth Factor Harvest Hematopoietic stem cells Histocytochemistry Human Immunodeficient Mouse Immunophenotyping Implant In Vitro Label Laboratories Laser Scanning Confocal Microscopy Life Literature MCAM gene Medical Modeling Monitor Mus Natural regeneration Obesity Organ Osteogenesis Outcome Pathway interactions Patients Physicians Population Process Regenerative Medicine Reporting Research Personnel Safety Semantics Silk Solid Sorting - Cell Movement Source Spleen Stem cell transplant Stem cells Surface Surface Antigens Testing Tissue Engineering Tissues Transgenes Transgenic Mice Transplantation adult stem cell base collagenase in vivo lipid biosynthesis mouse model progenitor public health relevance reconstitution regenerative research study scaffold self-renewal stem stem cell biology stem cell population stromal progenitor subcutaneous transcription factor

展开

项目摘要

DESCRIPTION (provided by applicant): Regenerative medicine and tissue engineering require an accessible, abundant, and reproducible source of cells that fulfill efficacy and safety requirements. During the past decade, basic science and clinical investigators have turned to adipose tissue as a cell source; however, there remains substantial controversy concerning the functional identity of the adipose-derived cells. Are they "stromal" or "stem" cells? This is not simply a semantic distinction. A true "stem" cell has the ability both to self-renew and to differentiate along multiple lineage pathways. While adipose-derived cells have been demonstrated to display these feature in vitro, the critical "gold standard" experiment, serial transplantation and reconstitution of a functional adipose depot in vivo, has never been reported in the literature. This revised R21 proposal tests our hypothesis that adipose tissue contains adipogenic "stem" cells in both the heterogeneous stromal vascular fraction (SVF) cells derived by collagenase digest of the adipose tissue and the relatively homogeneous, adherent, culture expanded adipose-derived stromal/stem cells (ASC). Aim 1 will employ the C57BL/6-Tg (UBC-GFP) 30cha/J mouse model as a source of easily trackable GFP+ adipose derived SVF cells or ASCs. Flow cytometry sorted cells will be seeded onto silk scaffolds and implanted into wild type C57BL/6 recipients. After a 4 week period, the constructs will be harvested and the process of transplantation repeated serially. Outcome parameters will be monitored and quantified by flow cytometry, histochemistry, confocal microscopy, scanning electron microscopy, and colony forming unit assays. Serial transplantation of intact adipose tissue explants will serve as a positive control. Aim 2 will perform comparable studies using flow cytometry sorted human SVF cells and ASC transplanted into immunodeficient mice. We anticipate that adipose tissue will prove to contain a stem cell population capable of reconstituting an intact fat pad upon serial transplantation.

描述（由申请人提供）：再生医学和组织工程需要满足功效和安全要求的可访问，丰富且可重复的细胞来源。在过去的十年中，基础科学和临床研究人员已将脂肪组织作为细胞来源。然而，关于脂肪衍生细胞的功能身份仍然存在重大争议。它们是“基质”或“茎”细胞？这不仅仅是语义上的区别。真正的“干式”细胞具有自我更新和沿多个谱系途径区分的能力。虽然已经证明脂肪衍生的细胞在体外显示了这些特征，但文献中从未报道过临界的“金标准”实验，体内功能性脂肪仓库的连续移植和重建。这项修订的R21提案检验了我们的假设，即脂肪组织在两个通过脂肪组织的胶原酶消化和相对均匀的，粘附的，粘附的，培养的脂肪蛋白培养的扩展的脂肪层依赖的脂肪组织中，均通过胶原酶消化产生的异质性基质血管分数（SVF）细胞中含有脂肪生成“干”细胞。 AIM 1将使用C57BL/6-TG（UBC-GFP）30CHA/J小鼠模型作为易于跟踪的GFP+脂肪衍生的SVF单元或ASC的来源。流式细胞术分类的细胞将被播种到丝绸支架上，并植入野生型C57BL/6受体中。经过4周的时间后，将收集构建体，并串行移植过程。结果参数将通过流式细胞仪，组织化学，共聚焦显微镜，扫描电子显微镜和菌落形成单元分析来监测和量化。完整脂肪组织外植体的连续移植将作为一个阳性对照。 AIM 2将使用流式细胞术分类的人类SVF细胞进行可比的研究，并将ASC移植到免疫缺陷的小鼠中。我们预计，脂肪组织将被证明包含能够在连续移植后重新建立完整脂肪垫的干细胞种群。