Jamur M C, Grodzki A C, Moreno A N, Swaim W D, Siraganian R P, Oliver C
Departamento de Biologia Celular, Universidade Federal do Paraná, Curitiba, PR, Brazil.
J Histochem Cytochem. 1997 Dec;45(12):1715-22. doi: 10.1177/002215549704501215.
Mast cells are difficult to purify from heterogeneous cell populations and to preserve, especially for pre-embedding immunostaining at the ultrastructural level. We have developed a technique that permits the isolation of a pure population of mast cells suitable for immunocytochemical studies. A rat mast cell-specific monoclonal antibody (MAb AA4) conjugated to tosylactivated Dynabeads 450 was used to immunomagnetically separate mast cells from rat bone marrow and peritoneal cell suspensions. Approximately 85% of the mast cells were recovered in the positive population that comprised virtually pure mast cells. After microwave fixation, morphological examination showed that the cells were intact and retained their ultrastructural detail. Mast cells in all stages of maturation were immunolabeled with a panel of antibodies after immunomagnetic separation. The combination of immunomagnetic separation followed by immunostaining should prove useful for the study of mast cell maturation and for the characterization of other specific cell types that are present in tissues in only limited numbers.
肥大细胞很难从异质性细胞群体中纯化出来并加以保存，尤其是在超微结构水平进行包埋前免疫染色时。我们开发了一种技术，可分离出适合免疫细胞化学研究的纯肥大细胞群体。将与甲苯磺酰活化的磁珠450偶联的大鼠肥大细胞特异性单克隆抗体（MAb AA4）用于从大鼠骨髓和腹腔细胞悬液中免疫磁分离肥大细胞。约85%的肥大细胞在包含几乎纯肥大细胞的阳性群体中被回收。微波固定后，形态学检查显示细胞完整并保留了其超微结构细节。免疫磁分离后，用一组抗体对所有成熟阶段的肥大细胞进行免疫标记。免疫磁分离后再进行免疫染色的组合应被证明对肥大细胞成熟的研究以及对仅以有限数量存在于组织中的其他特定细胞类型的表征有用。
