多发性骨髓瘤细胞系CZ-1的建立及其生物学特性

Establishment and biological characteristics of human multiple myeloma cell line CZ-1

目的 建立多发性骨髓瘤 (MM)细胞系并鉴定其生物学特性。方法 分离 1例晚期λ轻链型MM患者外周血和骨髓的单个核细胞 ,用液体培养法进行培养。用瑞特 姬姆萨染色和细胞化学染色确定细胞形态 ;用免疫固定电泳技术检测细胞是否分泌单克隆λ轻链 ,用荧光定量PCR法检测细胞是否被EB病毒感染 ;用流式细胞仪分析细胞的免疫表型 ;用染色体G分带法研究细胞的遗传学特征。结果 所得细胞系在饲养细胞或条件培养液存在时 ,可以持久增殖。该细胞分泌λ轻链 ,EB病毒阴性 ;细胞在瑞特 姬姆萨染色下呈典型的原、幼浆细胞形态 ;细胞化学染色 :酸性磷酸酶染色 (+) ,髓过氧化物酶 (- ) ;细胞免疫表型为CD1 0+,CD2 8+,CD3 8+,CD1 3 8+,CD56+,CD49d+,CD54+,CD44+,CD58+,而不表达CD1 9,CD40 ,CD95,CD95L ,CD3 4 ,CD2 ,CD5。骨髓来源和外周血来源的细胞系免疫表型无明显不同 ,核型分析结果均为i(1q +) ,8q +,13q +,i(17q) ,i(18q) ,+M。结论 用 1例MM患者的外周血和骨髓细胞同时建立了一个生物学特性相同的分泌λ轻链的MM细胞系CZ 1。

Objective To establish a multiple myeloma (MM) cell line and analyze its biological characteristics. Methods Mononuclear cells isolated from the peripheral blood(PB) and bone marrow(BM) of an advanced MM patient (λ light chain type) were incubated by liquid cell culture,cell morphology was analyzed by Wright-Giemsa-staining and cytochemical staining, immunophenotyping by flow cytometry, cytogenetic analysis by chromosome RHG-banding method.Quantitative fluorescent polymerase chain reaction was used to detect EBV DNA. Results The established cell line could survive and proliferate in the presence of feeder cells or conditioned medium. The cells secreted λ light chain and were negative for EBV. The Wright-Giemsa-staining showed typical plasmablast or plasma cell morphology. The cytochemical staining of the cells showed the following reactivity pattern:positive for acid phosphatase, negative for myeloperoxidase. The immunoprofile of the cells was concordant with that of MM cells: positive for CD 10,CD 28,CD 38,CD 138,CD 56,CD 49d,CD 44,CD 54 and CD 58,negative for CD 19,CD 40,CD 95,CD 95L,CD 34, CD 2 and CD 5. The cytogenetic analysis showed complex chromosome abnormality of i(1q+),8q+,13q+,i(17q),i(18q) and +M. There was no difference in morphology, immunophenotype and cytogenetics between cells from PB and BM. Conclusion A MM cell line secreting λ light chain named CZ-1,was established.The cells from both PB and BM have the same biological characteristics.