Moolten F L, Schreiber B
J Immunol Methods. 1980;36(3-4):359-68. doi: 10.1016/0022-1759(80)90141-6.
Murine thymus and spleen cells formed adherent monolayers in polystyrene tissue culture flasks when plated in serum-free medium. In the presence of 2% serum, thymus cells adhered poorly, but adherence was greatly enhanced if the flasks had been coated noncovalently with the lectin, concanavalin A. Adherence of leukemic lymphocytes (L1210) required both serum-free medium and concanavalin A-coated flasks; the extent of attachment was proportional to the concentration of the lectin used to coat the flasks at concentrations up to 0.1 mg/ml. Once L1210 cells had attached, they could not be removed by exposure to serum, ethylenediamine tetraacetic acid, trypsin, or alpha-methyl mannoside. Adherent L1210 cells remained capable of metabolism and proliferation during intervals of up to 7 days. The use of adherent monolayers for cytotoxicity assays was demonstrated by an assay for Pseudomonas aeruginosa toxin in EL4 murine leukemia cells.
当接种于无血清培养基中时，小鼠胸腺细胞和脾细胞能在聚苯乙烯组织培养瓶中形成贴壁单层。在含有2%血清的情况下，胸腺细胞贴壁不佳，但如果培养瓶用凝集素伴刀豆球蛋白A进行非共价包被，贴壁则会显著增强。白血病淋巴细胞（L1210）的贴壁既需要无血清培养基，也需要用伴刀豆球蛋白A包被的培养瓶；在高达0.1mg/ml的浓度范围内，附着程度与用于包被培养瓶的凝集素浓度成正比。一旦L1210细胞附着后，通过暴露于血清、乙二胺四乙酸、胰蛋白酶或α-甲基甘露糖苷都无法将其去除。贴壁的L1210细胞在长达7天的时间间隔内仍具有代谢和增殖能力。通过对EL4小鼠白血病细胞中的铜绿假单胞菌毒素进行检测，证明了贴壁单层用于细胞毒性检测的实用性。
