ROLE OF OSTEOBLASTS IN STEM CELL PROLIFERATION

成骨细胞在干细胞增殖中的作用

• 批准号: 6574772
• 负责人: STEPHEN G EMERSON
• 金额: $ 34.88万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-03-01 至 2003-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6574772
• 关键词: NOD mouse; SCID mouse; bone marrow; cell proliferation; cell transplantation; clinical research; connective tissue stroma; growth factor; hematopoiesis; hematopoietic stem cells; human subject; human tissue; laboratory mouse; mixed tissue /cell culture; osteoblasts; tissue /cell culture

Description (taken directly from the application): Hematopoietic stem cell survival, self-renewal and differentiation is believed to be highly dependent on the stromal microenvironment, which after birth is largely restricted to the bone marrow. However, investigations of candidate bone marrow stromal cells to date have concentrated on fairly ubiquitous cellular elements, such as fibroblasts and endothelial cells. As important as such cells may be in hematopoiesis, their activities cannot easily explain why hematopoiesis concentrates in the marrow. We therefore have recently begun to focus on a specific cell type, the osteoblast, which is located specifically on the endosteal surface within bone marrow cavities. Based upon our preliminary data, osteoblasts appear to directly communicate with primitive hematopoietic cells and support their proliferation. We propose to explore in depth the cellular and molecular basis for osteoblast-stem interactions. Specifically, we plan to: 1. Compare of the proliferative and self-renewal capacity of primitive hematopoietic cells before and after culture on osteoblast monolayers, utilizing: a. In vitro clonogenic and long term culture initiating cell assays; and b. In vivo transplantation assays using NOD/SCID xenogeneic transplant hosts, via limiting dilution and serial transplantation assays; 2. Determine the effect of bone marrow microenvironmental anatomy (adjacent hematopoietic cells) and physiology (perfusion) on osteoblast hematopoietic function; and 3. Molecularly clone osteoblast derived cDNAs whose protein products stimulate the proliferation of primitive human hematopoietic cells. These experiments should give a comprehensive picture of the hematopoietic role of an important but previously uncharacterized cell in the bone marrow microenvironment, a role which we hypothesize to be central to the maintenance and self-renewal of hematopoietic stem cells. Within the context of this Program Project application, these investigations will focus on the role and mechanisms by which marrow microenvironments govern stem cell self-renewal of the primitive stem cell compartments.

描述(直接取自应用程序)：造血干细胞 细胞存活， 自我更新和分化被认为高度依赖于 基质微环境，出生后主要局限于 骨髓。然而，对候选骨髓的调查 到目前为止，基质细胞主要集中在相当普遍的细胞上 元素，如成纤维细胞和内皮细胞。与 这样的细胞可能在造血，它们的活动不容易 解释为什么造血集中在骨髓中。因此，我们有 最近开始关注一种特定的细胞类型，成骨细胞，它是 位于骨髓内的骨内膜表面 龋齿。根据我们的初步数据，成骨细胞似乎 直接与原始造血细胞通信并支持其 扩散。我们建议深入探索细胞和分子 成骨细胞-干细胞相互作用的基础。具体来说，我们计划： 1.原始细胞增殖能力和自我更新能力的比较 成骨细胞单层培养前后的造血细胞 利用： A.体外克隆和长期培养启动细胞分析；以及 使用NOD/SCID异种移植进行体内移植试验 寄主，通过有限稀释和系列移植试验； 2.确定骨髓微环境解剖学的作用 (邻近造血细胞)和成骨细胞的生理(灌流) 造血功能；以及 3.分子克隆成骨细胞来源的cDNA，其蛋白产物为 刺激原始人类造血细胞的增殖。 这些实验应该会给出一个全面的图景 一种重要但以前未确定特征的细胞在 骨髓微环境，一个我们假设的角色 对造血干细胞的维持和自我更新至关重要。 在本计划项目申请的上下文中，这些 研究将集中在骨髓的作用和机制上 微环境控制原始干细胞的自我更新 牢房隔间。