Pauly J L, Han T
J Lab Clin Med. 1976 Nov;88(5):864-72.
A whole blood microculture assay is described for measuring lymphocyte reactivity to mitogenic and antigenic stimulants. This assay employs heparinized whole blood, serum-free culture medium, microtiter plates, and a Multiple Automated Sample Harvester (MASH). When this assay is compared to other leukocyte assays, its major advantages include (1) the utilization of fewer lymphocytes per microculture, thuus reducing the amount of blood required per test while increasing the number of test agents and replicate cultures which can be employed in any given experiment; (2) the conservation of mitogens, antigens, drugs, enzymes, hormones, lymphokines, and other test agents, some of which are either expensive of difficult to prepare in large quantities; (3) the elimination of lymphocyte isolation and purification procedures which may disrupt the relative proportion of T cells, B cells and antigen-processing cells; and (4) the application of an automated harvester which simplifies and expedites procedures required for processing cells for liquid scintillation counting.
本文描述了一种全血微量培养测定法,用于测量淋巴细胞对有丝分裂原和抗原刺激物的反应性。该测定法使用肝素化全血、无血清培养基、微量滴定板和多自动样品采集器(MASH)。当将该测定法与其他白细胞测定法进行比较时,其主要优点包括:(1)每个微量培养物使用的淋巴细胞较少,从而减少每次测试所需的血量,同时增加可用于任何给定实验的测试剂数量和重复培养次数;(2)节约有丝分裂原、抗原、药物、酶、激素、淋巴因子和其他测试剂,其中一些要么昂贵,要么难以大量制备;(3)无需进行淋巴细胞分离和纯化程序,这些程序可能会破坏T细胞、B细胞和抗原处理细胞的相对比例;(4)使用自动采集器,简化并加快了处理细胞进行液体闪烁计数所需的程序。