Gottshall S L, Hansen P J
Department of Dairy and Poultry Sciences, University of Florida, Gainesville 32611-0920.
Zentralbl Veterinarmed B. 1994 Oct;41(7-8):541-7. doi: 10.1111/j.1439-0450.1994.tb00261.x.
Conditions were established to optimize the lymphocyte proliferation test in sheep, a species where proliferation under standard conditions is lower than for other species. Optimum proliferation of phytohemagglutinin-stimulated lymphocytes was achieved in TCM-199 supplemented with 5% (v/v) horse serum, 0.2 mM sodium pyruvate and 5 x 10(-5) M beta-mercaptoethanol. Proliferative responses in this medium were 2.5-fold higher or more than when using a standard medium (RPMI-1640 + 10% heat-inactivated fetal bovine serum). Lymphocytes cultured in medium containing horse serum also responded to concanavalin A as well as or better than lymphocytes cultured in medium with bovine serum. For both PHA and Con A, optimal incorporation of radioactivity was achieved when cells were pulsed with [3H-methyl]-thymidine 72 h after addition of mitogen. The medium based on TCM-199 + horse serum could support interleukin-2-induced proliferation. Furthermore, known inhibitors of lymphocyte proliferation in sheep were effective in inhibiting lymphocytes cultured in this medium.
已建立条件以优化绵羊淋巴细胞增殖试验，在该物种中，标准条件下的增殖低于其他物种。在补充有5%（v/v）马血清、0.2 mM丙酮酸钠和5×10⁻⁵ Mβ-巯基乙醇的TCM-199中，可实现植物血凝素刺激的淋巴细胞的最佳增殖。在此培养基中的增殖反应比使用标准培养基（RPMI-1640 + 10%热灭活胎牛血清）时高2.5倍或更多。在含有马血清的培养基中培养的淋巴细胞对刀豆球蛋白A的反应与在含有牛血清的培养基中培养的淋巴细胞相同或更好。对于PHA和Con A，在添加促有丝分裂原72小时后用[³H-甲基]-胸腺嘧啶核苷脉冲细胞时，可实现放射性的最佳掺入。基于TCM-199 +马血清的培养基可支持白细胞介素-2诱导的增殖。此外，已知的绵羊淋巴细胞增殖抑制剂可有效抑制在此培养基中培养的淋巴细胞。
