Slide chamber culture system for the in vitro study of humoral regulation of granulocyte and monocyte-macrophage proliferation and differentiation
Slide chamber culture system for the in vitro study of humoral regulation of granulocyte and monocyte-macrophage proliferation and differentiation
复制标题
用于粒细胞和单核巨噬细胞增殖和分化的体液调节体外研究的载玻片室培养系统
DOI：
10.1182/blood.v50.2.289.289
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发表时间：
1977
期刊：
影响因子：
20.3
通讯作者：
Jm Jr Kinkade
中科院分区：
文献类型：
--
作者：
E. Winton;W. Vogler;K. Kellar;M. Parker;Jm Jr Kinkade
为了研究小鼠嗜中性粒细胞和单核-巨噬细胞系列细胞的增殖和分化，我们开发了一种采用载玻室的液体培养系统。用Ficoll-Hypaque上分离的1-2.5 X 10(4)个轻密度骨髓细胞开始培养。在存在集落刺激活性（CSAHU）时，观察到两种类型的簇。一个是紧密的球形，由中性粒细胞组成，而另一个是由单核-巨噬细胞系列的扁平细胞组成的松散组。玻片室培养适用于显微分析技术（如簇大小、放射自显影、免疫荧光）以及定量生化方法（如3h -胸腺嘧啶掺入率、DNA定量）。我们已经证明了紧密聚集的粒细胞的生成时间缩短和3H-胸腺嘧啶并入DNA的速率增加，随着CSAHU浓度的增加，总培养DNA也相应增加。通过组织化学染色和对中性粒细胞次级颗粒（乳铁蛋白）特异性标记蛋白的抗体的免疫荧光，证实了紧密球形团簇中的粒细胞分化。
A liquid culture system employing slide chambers was developed to facilitate the study of proliferation and differentiation of mouse neutrophilic granulocytes and cells of the monocyte-macrophage series. Cultures were initiated with 1–2.5 X 10(4) light density marrow cells which had been fractionated on Ficoll-Hypaque. In the presence of colony-stimulating activity (CSAHU), two types of clusters were observed. One was tight, spheroidal, and composed of neutrophilic granulocytes, while the other was a loose grouping of flattened cells of the monocyte-macrophage series. The slide chamber culture is adaptable to microscopic assay techniques (e.g., cluster size, autoradiography, immunofluorescence) as well as quantitative biochemical methods (e.g., rate of 3H-thymidine incorporation, DNA quantitation). We have demonstrated both a shortening of the generation time of granulocytes in tight clusters and increasing rates of 3H- thymidine incorporation into DNA, with a corresponding increase in total culture DNA as a function of CSAHU concentration. Granulocytic differentiation in the tight spheroidal clusters has been demonstrated by histochemical stains and immunofluorescence of an antibody to a marker protein specific for the secondary granule of the neutrophil (lactoferrin).