Tissue Reconstituting Potential of Human Stem Cells

人类干细胞的组织重建潜力

• 批准号: 7046860
• 负责人: Makio Ogawa
• 金额: $ 27.68万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7046860
• 关键词: NOD mouse; SCID mouse; cell differentiation; cell growth regulation; clinical research; cord blood; fluorescent in situ hybridization; hematopoietic stem cells; human subject; immunocytochemistry; newborn animals; polymerase chain reaction; xenotransplantation

DESCRIPTION (provided by applicant): Studies in rodent models suggest that bone marrow stem cells are capable of reconstituting a variety of tissues including not only mesenchymal tissues such as bone, cartilage, ligament, adipocytes, skeletal and cardiac muscles but also hepatocytes and CNS cells. This application pertains to studies of tissue reconstituting potential of human cord blood and adult stem cells based on a mouse/human xenograft model developed in our laboratory. In this model, "conditioned" newborn NOD/SCID/Beta2-microglobulin-null mice supported high-level hematopoietic engraftment by human cord blood cells. Recent studies using this model indicated that cord blood stem cells are capable of generating human hepatocytes and may also have the abilities to generate exocrine pancreatic cells and cardiomyocytes. This grant proposal is to determine the tissue reconstituting potential of human cord blood stem cells using this neonatal mouse xenograft model. Attempts will also be made to identify and purify the stem cells that are present in the cord blood by determining the surface phenotypes of tissue-engrafting cells. The major techniques to be used are fluorescence in situ hybridization (FISH) to identify human cells and immunohistochemistry, RT-PCR, and in situ hybridization for mRNA to delineate the tissue/cell lineages of the engrafted human cells. Comparison of these results with a separate study in our laboratory of tissue reconstituting potential of adult mouse stem cells indicates that there may be significant developmental control of the stem cell potentials. Based on these observations the following Specific Aims are proposed. Specific Aim #1: To determine the tissue reconstituting potential of human cord blood stem cells in a neonatal mouse xenograft model. Specific Aim #2: To identify and purify the human tissue reconstituting cells using the neonatal mouse xenograft model. Specific Aim #3: To determine the developmental control of the tissue reconstituting potentials of the stem cells in neonatal and adult mouse xenografts.

描述（申请人提供）：啮齿动物模型的研究表明，骨髓干细胞能够重建多种组织，不仅包括间充质组织，如骨、软骨、韧带、脂肪细胞、骨骼和心肌，还包括肝细胞和中枢神经系统细胞。该应用涉及基于我们实验室开发的小鼠/人类异种移植模型的人类脐带血和成体干细胞的组织重建潜力的研究。在该模型中，“条件化”新生NOD/SCID/Beta2-微球蛋白缺失小鼠支持人类脐带血细胞的高水平造血植入。最近使用该模型的研究表明，脐带血干细胞能够产生人类肝细胞，并且还可能具有产生外分泌胰腺细胞和心肌细胞的能力。该拨款提案旨在利用新生小鼠异种移植模型确定人脐带血干细胞的组织重建潜力。还将尝试通过确定组织移植细胞的表面表型来鉴定和纯化脐带血中存在的干细胞。使用的主要技术是荧光原位杂交 (FISH) 来鉴定人类细胞，以及免疫组织化学、RT-PCR 和 mRNA 原位杂交来描绘移植人类细胞的组织/细胞谱系。将这些结果与我们实验室对成年小鼠干细胞的组织重建潜力的单独研究进行比较表明，干细胞潜力可能存在显着的发育控制。根据这些观察，提出以下具体目标。 具体目标#1：确定人脐带血干细胞在新生小鼠异种移植模型中的组织重建潜力。 具体目标#2：使用新生小鼠异种移植模型鉴定和纯化人体组织重建细胞。 具体目标#3：确定新生和成年小鼠异种移植物中干细胞组织重建潜力的发育控制。