Musashi M
Third Department of Internal Medicine, Hokkaido University School of Medicine, Sapporo, Japan.
Hokkaido Igaku Zasshi. 1989 Sep;64(5):567-79.
Human peripheral blood mononuclear cells (PBMC) obtained from healthy volunteers gave rise to three types of colonies that could be distinguished by their unique morphological characteristics; 5.1 +/- 0.9 (mean +/- SE) loose colonies of small cells, 2.1 +/- 0.9 packed colonies of larger cells, and 1.6 +/- 0.7 mixed colonies, were formed when PBMC (2 x 10(5) cells/dish) from 10 healthy volunteers were cultured in the medium containing methylcellulose. Cytochemical analysis with Astra blue-eosin dual staining revealed that all types of colonies consisted of various proportions of basophils, eosinophils, and hybrid (eosinophilic and basophilic) granulocytes which contained both of granules. These hybrid granulocytes were also identified by the ultrastructural features of two kinds of the granules. Relationships between cell numbers added to culture and formed colony numbers indicated colony of the cells to form the colonies. The colonies formed from untreated patients with chronic myelogenous leukemia (CML) during chronic phase were sevenfold of those from healthy volunteers. The colonies formed from treated patients with CML were normal in number. The number was 40 times greater in culture from a patient with basophilic crisis and a patient with myeloid crisis than normal, whereas that from a patient with lymphoid crisis were within normal limit. The number of the colonies from PBMC of patients with eosinophilia were in normal range, whereas those from bone marrow were six times or more than those from PBMC. These findings suggest that PBMC contains common basophil-eosinophil progenitors, and the culture used in this study in considered to be useful in the examination of basophil and eosinophil production from PBMC. Further studies using more purified cell population and other sources of colony stimulating factors such as interleukin-3 (IL-3), IL-4, IL-5, and granulocyte-macrophage colony stimulating factor should be carried out in order to clarify the significance of hybrid granulocytes in basophil and eosinophil proliferation and differentiation.
从健康志愿者获取的人外周血单个核细胞(PBMC)可形成三种类型的集落,这些集落可通过其独特的形态特征加以区分;当将来自10名健康志愿者的PBMC(2×10⁵个细胞/培养皿)在含甲基纤维素的培养基中培养时,形成了5.1±0.9(均值±标准误)个小细胞的松散集落、2.1±0.9个较大细胞的紧密集落以及1.6±0.7个混合集落。用天青蓝 - 伊红双重染色进行细胞化学分析表明,所有类型的集落均由不同比例的嗜碱性粒细胞、嗜酸性粒细胞以及同时含有两种颗粒的杂交(嗜酸性和嗜碱性)粒细胞组成。这些杂交粒细胞也可通过两种颗粒的超微结构特征得以鉴定。添加到培养物中的细胞数量与形成的集落数量之间的关系表明了形成集落的细胞的集落情况。慢性期未经治疗的慢性粒细胞白血病(CML)患者形成的集落是健康志愿者的7倍。接受治疗的CML患者形成的集落数量正常。嗜碱性危象患者和髓细胞危象患者培养物中的集落数量比正常情况大40倍,而淋巴细胞危象患者的集落数量在正常范围内。嗜酸性粒细胞增多症患者PBMC形成的集落数量在正常范围内,而骨髓来源的集落数量是PBMC来源的6倍或更多。这些发现提示PBMC含有常见的嗜碱性粒细胞 - 嗜酸性粒细胞祖细胞,并且本研究中使用的培养方法被认为有助于检测PBMC中嗜碱性粒细胞和嗜酸性粒细胞的生成。为了阐明杂交粒细胞在嗜碱性粒细胞和嗜酸性粒细胞增殖与分化中的意义,应使用更纯化的细胞群体以及其他集落刺激因子来源,如白细胞介素 - 3(IL - 3)、IL - 4、IL - 5和粒细胞 - 巨噬细胞集落刺激因子进行进一步研究。