De Vis J, Renmans W, Segers E, Jochmans K, De Waele M
Department of Hematology, Academic Hospital, Free University of Brussels, (V.U.B.), Belgium.
J Immunol Methods. 1991 Mar 21;137(2):193-7. doi: 10.1016/0022-1759(91)90024-a.
Immunophenotyping of leukemia cells is generally performed on cells in suspension. These suspensions are usually prepared from anticoagulated peripheral blood or bone marrow samples but when anticoagulation is suboptimal clotted samples may reach the laboratory. In this study cell suspensions were prepared from clotted blood and bone marrow samples of leukemia patients by lysis of the clots with streptokinase. The cell suspensions were then labeled with monoclonal or polyclonal antibodies and examined by flow cytometry or fluorescence microscopy. Enough cells could be isolated from small volumes of clotted blood or bone marrow aspirate to determine the immunophenotype of the cells. The morphology of the cells was well preserved and accurate identification of the cells was possible. The immunophenotypes determined on clotted samples from four patients with acute myeloblastic leukemia, five with acute lymphoblastic leukemia and two with chronic lymphocytic leukemia were identical to those established using EDTA anticoagulated samples of the same patients. When no unclotted samples are available this approach may avoid the necessity for a new sample and the concomitant delay in treatment of the patient.
白血病细胞的免疫表型分析通常在悬浮细胞上进行。这些悬浮液通常由抗凝外周血或骨髓样本制备，但当抗凝效果不佳时，凝血样本可能会送到实验室。在本研究中，通过用链激酶溶解凝血块，从白血病患者的凝血血液和骨髓样本中制备细胞悬浮液。然后用单克隆或多克隆抗体标记细胞悬浮液，并通过流式细胞术或荧光显微镜检查。从小体积的凝血血液或骨髓抽吸物中可以分离出足够的细胞来确定细胞的免疫表型。细胞形态保存良好，能够准确识别细胞。对4例急性髓细胞白血病、5例急性淋巴细胞白血病和2例慢性淋巴细胞白血病患者的凝血样本进行的免疫表型分析与使用同一患者的乙二胺四乙酸（EDTA）抗凝样本所确定的免疫表型相同。当没有未凝血样本时，这种方法可以避免重新采样的必要性以及随之而来的患者治疗延迟。
