CORE RESOURCE: BLOOD AND BONE MARROW COLECTION CORE

核心资源：血液和骨髓采集核心

• 批准号: 7376281
• 负责人: DARWIN Johnson PROCKOP
• 金额: $ 4.8万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-12-01 至 2006-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7376281
• 关键词: CORE; RESOURCE; BLOOD; BONE; MARROW

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The cells have been variously referred to as colony-forming fibroblasts, mesenchymal stem cells, or marrow stromal cells (MSCs), and have been attracting increasing interest both for their biological properties as multipotential stem-like cells and their potential use for cell and gene therapy. However, a major problem in the field has been that reproducible conditions for isolation and expansion of the cells in culture has not been defined. The overall aim of the present proposal is to develop effective conditions for their isolation and expansion of human hMSCs in culture based on three recent observations made in our laboratory: 1) we have developed culture conditions whereby hMSCs can be expanded over 108-fold without significant loss in replicative capacity; 2) we have found that the replication of hMSCs in culture is regulated autocrine/paracrine factors that can be recovered from the culture medium; 3) we have identified a sub-population of small granular cells in cultures of hMSCs that are highly replicative and appear to be the earliest progenitors in the cultures. The specific aims of this protocol are: l) to determine whether the highly replicative cells we have obtained after 108-fold expansion of hMSCs retain their multiopotentiality to differentiate into osteoblasts, chondrocytes, and adipocytes; 2) isolate and characterize the secreted factors that regulate replication of hMSCs in culture; 3) isolate the small, granular, and highly replicative cells we have identified in cultures of hMSCs and define their surface epitopes.

该子项目是利用 NIH/NCRR 资助的中心拨款提供的资源的众多研究子项目之一。子项目和研究者 (PI) 可能已从另一个 NIH 来源获得主要资金，因此可以在其他 CRISP 条目中出现。列出的机构是中心的机构，不一定是研究者的机构。这些细胞被不同地称为集落形成成纤维细胞、间充质干细胞或骨髓基质细胞（MSC），并且因其作为多能干细胞样细胞的生物学特性及其在细胞和基因方面的潜在用途而引起了越来越多的兴趣。治疗。然而，该领域的一个主要问题是培养细胞分离和扩增的可重复条件尚未确定。本提案的总体目标是根据我们实验室最近的三项观察结果，开发在培养物中分离和扩增人类 hMSC 的有效条件：1）我们开发了培养条件，使 hMSC 可以扩增 108 倍以上，且不会产生明显的影响。复制能力丧失； 2）我们发现hMSC在培养物中的复制受到可从培养基中回收的自分泌/旁分泌因子的调节； 3) 我们在 hMSC 培养物中鉴定出了一个小颗粒细胞亚群，它们具有高度复制性，并且似乎是培养物中最早的祖细胞。该协议的具体目标是： l) 确定 hMSC 扩增 108 倍后获得的高度复制细胞是否保留其分化为成骨细胞、软骨细胞和脂肪细胞的多向潜能； 2) 分离并表征调节培养物中 hMSC 复制的分泌因子； 3) 分离我们在 hMSC 培养物中鉴定出的小颗粒状且高度复制的细胞，并确定其表面表位。