Crucian B E, Sams C F
Wyle Laboratories, Houston, TX, USA.
J Gravit Physiol. 1999 Jul;6(1):P33-4.
Significant changes have recently been described regarding circulating peripheral immune cells immediately following spaceflight. Existing methods for immunophenotype staining of peripheral blood in terrestrial labs do not meet the constraints for flight on the Space Shuttle. We have recently described the development and use of the Whole Blood Staining Device (WBSD), a simple device for staining flow cytometry specimens during spaceflight. When preparing samples with the WBSD, all liquids are safely contained as the cells are moved through staining, lysis and fixation steps. Here we briefly review the use of the WBSD, and then describe another versatile adaptation, a modification to perform intracellular staining of cytokines for detection by flow cytometry. Alterations in cytokine production have been reported both in ground-based simulated microgravity culture and in astronaut samples returning from spaceflight. Data regarding microgravity effects on cytokine production for specific subpopulations of cells is lacking. Flow cytometric cytokine analysis offers the unique ability to perform simultaneous surface marker analysis and positively identity cytokine producing subsets of cells. The utilization of the WBSD provides the ability to perform rapid and routine mitogenic activation during spaceflight coupled with the ability to perform simultaneous surface marker analysis. The only external requirements for this procedure are an in-flight 37-degree incubator and the capacity for 4-degree storage.
最近有关于太空飞行后循环外周免疫细胞的显著变化的描述。地面实验室中现有的外周血免疫表型染色方法不符合航天飞机飞行的限制条件。我们最近描述了全血染色装置（WBSD）的开发和使用，这是一种在太空飞行期间用于染色流式细胞术标本的简单装置。使用WBSD制备样品时，当细胞经过染色、裂解和固定步骤时，所有液体都能安全容纳。在此我们简要回顾WBSD的使用，然后描述另一种通用的改进方法，即对用于通过流式细胞术检测细胞因子的细胞内染色进行的改进。在地面模拟微重力培养和从太空飞行返回的宇航员样本中均已报道细胞因子产生的变化。缺乏关于微重力对特定细胞亚群细胞因子产生影响的数据。流式细胞术细胞因子分析具有同时进行表面标志物分析并准确鉴定产生细胞因子的细胞亚群的独特能力。WBSD的使用使得在太空飞行期间能够进行快速且常规的促有丝分裂激活，同时具备进行表面标志物分析的能力。此程序唯一的外部要求是一个飞行中的37度培养箱和4度储存的能力。
