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

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可获取、丰富且可重复的细胞来源，以满足功效和安全性要求。在过去的十年中，基础科学和临床研究人员已转向脂肪组织作为细胞来源。然而，关于脂肪源细胞的功能特性仍然存在很大争议。它们是“基质”细胞还是“干”细胞？这不仅仅是语义上的区别。真正的“干”细胞具有自我更新和沿多种谱系途径分化的能力。虽然脂肪来源的细胞已被证明在体外显示出这些特征，但关键的“金标准”实验，即体内功能性脂肪库的连续移植和重建，从未在文献中报道过。这项修订后的 R21 提案测试了我们的假设，即脂肪组织在通过脂肪组织的胶原酶消化衍生的异质基质血管部分 (SVF) 细胞和相对均质、贴壁、培养扩增的脂肪衍生基质/干细胞中均含有脂肪形成“干”细胞细胞（ASC）。目标 1 将采用 C57BL/6-Tg (UBC-GFP) 30cha/J 小鼠模型作为易于追踪的 GFP+ 脂肪来源的 SVF 细胞或 ASC 的来源。流式细胞仪分选的细胞将被接种到丝支架上并植入野生型 C57BL/6 受体中。 4周后，将收获构建体并连续重复移植过程。结果参数将通过流式细胞术、组织化学、共聚焦显微镜、扫描电子显微镜和集落形成单位测定进行监测和量化。完整脂肪组织外植体的连续移植将作为阳性对照。目标 2 将使用流式细胞仪分选的人类 SVF 细胞和移植到免疫缺陷小鼠体内的 ASC 进行类似研究。我们预计脂肪组织将被证明含有能够在连续移植后重建完整脂肪垫的干细胞群。