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用于生长因子依赖性和非依赖性浆细胞瘤及杂交瘤的无血清培养基。

Serum-free medium for growth factor-dependent and -independent plasmacytomas and hybridomas.

作者信息

Shacter E

出版信息

J Immunol Methods. 1987 May 20;99(2):259-70. doi: 10.1016/0022-1759(87)90136-0.

Abstract

A serum-free medium was developed for culture of plasmacytomas and hybridomas that are dependent upon or independent of the presence of specific P388D1-derived polypeptide growth factors. Possibly due to the stringency of requirements for culturing such plasmacytomas, a highly advantageous combination of components was developed. The medium, SFM, contains RPMI 1640, beta-mercaptoethanol, Hepes buffer, reduced glutathione (GSH), selenite (Se), pyruvate, transferrin, albumin, soybean lipids, and low density lipoproteins. A cooperative effect of GSH and Se is observed. SFM supports the continuous, prodigious growth of every B cell line tested, including a variety of the plasmacytomas, hybridomas, myeloma fusion partners and lymphoma cell lines, as well as the macrophage-like cell line P388D1. All of the B cell lines grow equally as well in SFM as in serum-containing medium. Hybridoma lines from mouse-mouse and rat-mouse fusions continue to produce monoclonal antibodies which can then be purified in a single-step procedure. The cells can be cloned out and frozen down in SFM. In addition, the medium is highly effective for establishment of plasmacytoma cell lines from some transplantable plasmacytomas, including early generation tumors.

摘要

已开发出一种无血清培养基,用于培养依赖或不依赖特定P388D1衍生多肽生长因子的浆细胞瘤和杂交瘤。可能由于培养此类浆细胞瘤的要求严格,已开发出一种极具优势的成分组合。该培养基SFM含有RPMI 1640、β-巯基乙醇、Hepes缓冲液、还原型谷胱甘肽(GSH)、亚硒酸盐(Se)、丙酮酸盐、转铁蛋白、白蛋白、大豆脂质和低密度脂蛋白。观察到GSH和Se的协同作用。SFM支持所测试的每一种B细胞系持续、大量生长,包括多种浆细胞瘤、杂交瘤、骨髓瘤融合伙伴和淋巴瘤细胞系,以及巨噬细胞样细胞系P388D1。所有B细胞系在SFM中的生长情况与在含血清培养基中一样好。来自小鼠-小鼠和大鼠-小鼠融合的杂交瘤细胞系继续产生单克隆抗体,然后可通过一步法进行纯化。细胞可在SFM中克隆并冻存。此外,该培养基对于从一些可移植浆细胞瘤(包括早期肿瘤)建立浆细胞瘤细胞系非常有效。

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