THE GROWTH-FACTOR REQUIREMENTS OF STRO-1-POSITIVE HUMAN BONE-MARROW STROMAL PRECURSORS UNDER SERUM-DEPRIVED CONDITIONS IN-VITRO
THE GROWTH-FACTOR REQUIREMENTS OF STRO-1-POSITIVE HUMAN BONE-MARROW STROMAL PRECURSORS UNDER SERUM-DEPRIVED CONDITIONS IN-VITRO
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DOI：
10.1182/blood.v85.4.929.bloodjournal854929
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发表时间：
1995-02-15
期刊：
影响因子：
20.3
通讯作者：
SIMMONS, PJ
中科院分区：
文献类型：
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作者：
GRONTHOS, S;SIMMONS, PJ
调节原始骨髓基质细胞前体细胞生长和发育的因素尚未明确。我们已经检查了25种纯化的重组生长因子启动和支持成人骨髓成纤维细胞集落形成细胞（CFU-F）克隆形成生长的能力。使用单克隆抗体（MoAb）STRO-1，通过磁激活细胞分选（MACS），使用CFU-F富集的骨髓单核细胞（BMMNC）进行试验。开发了一种血清去除试验，以避免胎牛血清（FCS）中可能掩盖或以其他方式改变CFU-F对外源性添加因子的反应的成分。L-抗坏血酸和糖皮质激素地塞米松被认为是必需的CFU-F集落发育在血清剥夺条件下。重要的是，CFU-F在该培养系统中的克隆形成性生长完全依赖于生长因子的外源性来源。血小板衍生生长因子-BE（PDGF）和表皮生长因子（EGF）表现出最大的支持集落生长的能力。菌落形成呈剂量依赖性，任一因子的半数最大菌落数约为0.2 ng/mL，浓度超过1.0 ng/mL时达到平台数。同时添加PDGF和EGF对启动的集落数量没有影响，但与单独使用任一因子或20%FCS在对照培养物中诱导的集落大小相比，平均集落直径显著（P小于或等于0.05）呈剂量依赖性增加。使用针对PDGF受体α链和EGF受体α链的MoAb与Moab STR 0 -1组合进行的BMMNC的趋化激活细胞分选（FAGS）证实了两种受体在CFU-F上的组成型表达大于90%。在STRO-1（+）CFU-F上也检测到胰岛素样生长因子-1（IGF-1）和神经生长因子（NGF）的受体，但在体外，IGF-1和NGF均不支持集落生长。本报告证明了一个简单的，可重复的，严格的培养系统的发展，在血清剥夺条件下的基质前体的生长和测定，并代表了一个重要的先决条件，为未来的研究中的作用，生长因子在调节基质细胞增殖，分化和发展。(C)1995年，美国血液学会。
Factors that regulate the growth and development of primitive bone marrow stromal cell precursors are not well defined. We have examined 25 purified recombinant growth factors for their ability to initiate and support clonogenic growth of fibroblast colony-forming cells (CFU-F) from adult human bone marrow. Assays were performed using bone marrow mononuclear cells (BMMNC) enriched in CFU-F by magnetic-activated cell sorting (MACS) using the monoclonal antibody (MoAb) STRO-1. A serum-deprived assay was developed to avoid components of fetal calf serum (FCS) that may mask or otherwise modify the response of CFU-F to exogenously added factors. L-ascorbate and the glucocorticoid dexamethasone were found to be essential for CFU-F colony development under serum-deprived conditions. Importantly, clonogenic growth of CFU-F in this culture system was absolutely dependent on an exogenous source of growth factor. Platelet-derived growth factor-BE (PDGF) and epidermal growth factor (EGF) demonstrated the greatest ability to support colony growth. Colony formation was dose-dependent, with half-maximal colony numbers at approximately 0.2 ng/mL for either factor and plateau numbers at concentrations in excess of 1.0 ng/mL. Simultaneous addition of PDGF and EGF had no effect on the number of colonies initiated but resulted in dose-dependent increases in mean colony diameter that were significant (P less than or equal to .05) when compared with the effect of either factor alone or with the size of colonies elicited in control cultures by 20% FCS. Fluorescence-activated cell sorting (FAGS) of BMMNC using MoAbs to the alpha chain of the PDGF receptor and to the EGF receptor in combination with the Moab STRO-1 demonstrated constitutive expression of both receptors by greater than 90% on CFU-F. Receptors for insulin-like growth factor-1 (IGF-1) and nerve growth factor (NGF) were also detected on STRO-1(+) CFU-F, but in vitro both IGF-1 and NGF did not support colony growth. This report demonstrates the development of a simple, reproducible, and stringent culture system for the growth and assay of stromal precursors under serum-deprived conditions and represents an important prerequisite for future studies of the role of growth factors in the regulation of stromal cell proliferation, differentiation, and development. (C) 1995 by The American Society of Hematology.