长期培养人急性髓系白血病骨髓基质上清液对早幼粒白血病细胞的影响  

Influence of long-term cultured acute myeloid leukemia stroma supernatant on acute promyelocytic leukemia cells

作　　者：孙健[1] 宋振岚[1] 

机构地区：[1]大连医科大学附属第二医院血液内科,辽宁省大连市116027

出　　处：《中国组织工程研究与临床康复》2007年第15期2893-2896,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：目的:通过观察长期培养的白血病骨髓基质上清液对早幼粒白血病细胞体外的作用,寻求一种体外净化辅助方法。方法:实验于2002-04/2003-12在大连医科大学附属第二医院血液内科实验室完成。20例急性髓细胞白血病为初诊或复发病例。男11例,女9例,19～55岁,平均36.8岁,由大连医科大学附属第二医院血液内科、大连市友谊医院血液内科、铁路医院血液科、大连大学附属医院血液科提供。患者知情同意并签署知情同意书。诊断均符合张之南主编的“血液病诊断及疗效标准”,其中M12例,M28例,M34例,M42例,M54例。常规分离培养人骨髓基质细胞,收集14,28,35d的上清分别与早幼粒白血病细胞共孵育,以不加入急性髓细胞白血病基质上清液的早幼粒白血病细胞的培养体系作为阴性对照,通过形态、流式细胞术、DNA电泳来观察早幼粒白血病细胞有无分化及凋亡表现。结果:20例患者均进入结果分析。①急性髓细胞白血病骨髓基质细胞是由网状细胞、成纤维细胞、巨噬细胞、脂肪细胞组成。②Wright-Giemsa染色可见阴性对照早幼粒白血病细胞为悬浮生长,胞体内含颗粒,增殖旺盛,镜下常见核分裂、增殖状态的早幼粒白血病细胞;14d及28d培养的急性髓细胞白血病基质上清液作用的早幼粒白血病细胞,形态未见明显变化,35d培养的急性髓细胞白血病基质上清液作用的早幼粒白血病细胞出现细胞变形,细胞核出现改变,部分细胞核仁减少或消失。NBT还原实验发现部分早幼粒白血病细胞胞奖内出现紫蓝色颗粒,并见到核碎解成染色质小体(凋亡小体)。③35d时相上清液孵育的早幼粒白血病细胞部分出现分化现象,部分出现凋亡小体,DNA电泳见梯形带,流式细胞术检测出凋亡峰。④培养35d的急性髓细胞白血病基质上清液对早幼粒白血病细胞的诱导分化及促凋亡作用明显强于培养14,28d的急性髓细�AIM： Trough observing the effect of the supemates from long-term cultured Acute Myeloid Leukemia （AML） stroma cells on the acute promyelocytic leukemia cells, to find an assistant purging method in vitro. METHODS： The experiment was performed at the Laboratory of Department of Hematology, Second Affiliated Hospital, Dalian Medical University from April 2002 to December 2003. The 20 cases of AML were preliminarily diagnosed or reoccurrence, including 11 males and 9 females, aged from 19-55 years, averagely 36.8 years. The cases were provided by Department of Hematology of Second Affiliated Hospital of Dalian Medical University, Department of Hematology of Dalian Friendship Hospital, Department of Hematology of Railway Hospital, and Department of Hematology of Affiliated Hospital of Dalian University. The patients knew the fact and agreed and signed the informed consent. The diagnosis met the Standard of Hematology Diagnoses and Therapy compiled by Zhang chiefly. There were 2 cases of M1, 8 cases of M2, 4 cases of M3, 2 cases of M4 and 4 cases of M5. AML stroma cells were isolated and cultured routinely. Supematant was collected at days 14, 28 and 35, and incubated with acute promyelocytic leukemia cells together. The culture medium of acute promyelocytic leukemia cells without AML stroma supernatant was used as negative control. Differentiation and apoptosis of acute promyelocytic leukemia cells were observed by form, flow cytometry, DNA electrophoresis. RESULTS： The 20 patients were involved in the result analysis. ①The AML stroma cells were composed of four kinds of cells： fibroblast, reticular cell, macrophage and lipocyte. ②Wright-Giemsa staining could found that the acute promyelocytic leukemia cells lived by the mode of suspending, which could be observed nuclear split and bloom proliferation through microscope in the negative control group. Acute promyelocytic leukemia cells cultured with stromal supernatant for 14 and 28 days did not change obviously. Those cultured for 35 days appeared

关 键 词：基质细胞 上清液 人早幼粒白血病细胞 体外净化 

分 类 号：R733.71[医药卫生—肿瘤]