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一种新的人骨髓瘤细胞系(U - 2030)的建立及对次黄嘌呤氨基蝶呤胸腺嘧啶核苷(HAT)敏感亚系的筛选

Establishment of a new human myeloma cell line (U-2030) and selection of a hat-sensitive subline.

作者信息

Jernberg H, Björklund G, Nilsson K

出版信息

Int J Cancer. 1987 Jun 15;39(6):745-51. doi: 10.1002/ijc.2910390615.

Abstract

We have established a new human myeloma cell line from the pleural effusion of a patient with an IgA lambda myeloma, using special tissue culture conditions and selection procedures to prevent the outgrowth of contaminating Epstein-Barr virus (EBV)-carrying normal B-lymphoblastoid cells present in the explant. The myeloma cell line, U-2030, is aneuploid and EBNA-negative and has morphological features, reactivity with cytochemical markers and cell-surface antigen expression typical of plasmablasts. The cell line thus appears to be representative of the malignant clone in vivo. However, functionally the line is a non-Ig-producer and must therefore be derived from a non-secretory variant cell present within the highly aneuploid myeloma cell clone in vivo. The U-2030 differs from previously established human myeloma cell lines in that it has a comparatively high growth rate, is clonable and can be made HAT-sensitive relatively easily. This, together with the facts that it is a non-Ig-producer and mycoplasma-free, suggests that the 6-thioguanine-resistant, HAT-sensitive subline, U-2030 TG, derived from this cell line may be used as a malignant fusion partner for the production of human-human hybridomas. An EBV-carrying lymphoblastoid cell line (U-2031) was also established. This line was diploid and had all the phenotypic properties of lymphoblastoid lines established from normal individuals.

摘要

我们从一名患有IgA λ型骨髓瘤患者的胸腔积液中建立了一种新的人骨髓瘤细胞系,采用特殊的组织培养条件和筛选程序,以防止外植体中携带爱泼斯坦-巴尔病毒(EBV)的正常B淋巴母细胞过度生长。骨髓瘤细胞系U-2030为非整倍体且EBNA阴性,具有成浆细胞典型的形态特征、与细胞化学标志物的反应性及细胞表面抗原表达。因此,该细胞系似乎代表了体内的恶性克隆。然而,从功能上来说,该细胞系不产生免疫球蛋白(Ig),因此必定来源于体内高度非整倍体骨髓瘤细胞克隆中的非分泌型变异细胞。U-2030与先前建立的人骨髓瘤细胞系不同,它具有相对较高的生长速率,可克隆,并且相对容易对氨基蝶呤-氨甲蝶呤-胸腺嘧啶核苷(HAT)敏感。这一点,再加上它不产生Ig且无支原体污染的事实,表明从该细胞系衍生出的对6-硫鸟嘌呤耐药、对HAT敏感的亚系U-2030 TG,可用作生产人-人杂交瘤的恶性融合伙伴。还建立了一个携带EBV的淋巴母细胞系(U-2031)。该细胞系为二倍体,具有从正常个体建立的淋巴母细胞系的所有表型特征。

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