Mattes M J, Tanimoto M, Pollack M S, Maurer D H
J Immunol Methods. 1983 Jul 15;61(2):145-50. doi: 10.1016/0022-1759(83)90156-4.
A simple method is described for preparing monolayers of non-adherent cells, using concanavalin A to bind the cells to wells of plastic microtest plates. The method was used successfully with all 202 human cell types tested, which included 23 tissue culture lines, 165 fresh specimens of all major histological types of leukemia and lymphoma, 20 fresh myelomas, 2 fresh thymomas, normal spleen and lymph node cells, fractionated T lymphocytes and B lymphocytes from peripheral blood, and cultured fetal amniotic cells. All cell types attached firmly, and were not detached by subsequent vigorous washing. In contrast, attempted attachment of cells in serum free medium, or with poly-L-lysine or glutaraldehyde, was ineffective with many cell types. We used the monolayers as target cells for antibodies to cell surface antigens, utilizing immune rosetting or complement-mediated cytotoxicity. This procedure should simplify most assays involving non-adherent target cells.
本文描述了一种制备非贴壁细胞单层的简单方法，该方法利用伴刀豆球蛋白A将细胞结合到塑料微量测试板的孔中。该方法成功应用于所测试的所有202种人类细胞类型，包括23种组织培养细胞系、165份所有主要组织学类型白血病和淋巴瘤的新鲜标本、20份新鲜骨髓瘤、2份新鲜胸腺瘤、正常脾细胞和淋巴结细胞、外周血中分选的T淋巴细胞和B淋巴细胞，以及培养的胎儿羊膜细胞。所有细胞类型均牢固附着，且在随后的剧烈洗涤中不会脱落。相比之下，在无血清培养基中，或使用聚-L-赖氨酸或戊二醛尝试使细胞附着，对许多细胞类型均无效。我们将这些单层细胞用作针对细胞表面抗原抗体的靶细胞，采用免疫玫瑰花结试验或补体介导的细胞毒性试验。该方法应能简化大多数涉及非贴壁靶细胞的检测。
