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，出生后很大程度上限于 骨髓。然而，对候选骨髓的研究 迄今为止，基质细胞主要集中在相当普遍的细胞上 元素，例如成纤维细胞和内皮细胞。一样重要 此类细胞可能处于造血功能，其活动不易 解释为什么造血作用集中在骨髓中。因此我们有 最近开始关注一种特定的细胞类型，即成骨细胞， 专门位于骨髓内的骨内膜表面 空洞。根据我们的初步数据，成骨细胞似乎 直接与原始造血细胞沟通并支持其 增殖。我们建议深入探索细胞和分子 成骨细胞-干相互作用的基础。具体来说，我们计划： 1. 原始细胞增殖和自我更新能力比较 在成骨细胞单层上培养之前和之后的造血细胞， 利用： 一个。体外克隆形成和长期培养起始细胞测定；和 b.使用 NOD/SCID 异种移植进行体内移植测定 宿主，通过有限稀释和连续移植测定； 2. 确定骨髓微环境解剖的影响 成骨细胞（邻近造血细胞）和生理学（灌注） 造血功能；和 3. 分子克隆成骨细胞衍生的 cDNA，其蛋白质产物 刺激人类原始造血细胞的增殖。 这些实验应该能够全面地描述 一个重要但以前未表征的细胞的造血作用 骨髓微环境，我们假设的作用是 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背景下，这些 研究将集中于骨髓的作用和机制 微环境控制原始干细胞的自我更新 细胞室。