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豚鼠淋巴细胞在含有和不含有巯基乙醇及胎牛血清的培养基中对有丝分裂原、一种抗原和混合淋巴细胞培养的反应。

Responses of guinea-pig lymphocytes to mitogens, an antigen, and mixed leucocyte culture in media with and without mercaptoethanol and foetal calf serum.

作者信息

Gregerson D S, Kelly B, Levy J G

出版信息

Immunology. 1975 Aug;29(2):237-46.

Abstract

The ability of guinea-pig spleen and lymph node cells to undergo a proliferative response in vitro in the presence of mitogens (concanavalin A and lipopolysaccharide), a specific antigen (oxidized ferredoxin), and allogeneic cells was assessed under a variety of conditions. Time and dose dependency of the responses was measured in RPMI 1640, RPMI 1640 plus mercaptoethanol (ME), RPMI 1640 plus foetal calf serum (FCS), and RPMI 1640 with ME and FCS. Mitogen responses were also measured after treatment of the cells with sheep antiguinea pig immunoglobulin (SaGPIg) and complement (C') or after passage through nylon wool columns. Lipopolysaccharide (LPS) stimulated the cells under all media conditions over a wide range of concentrations but over a narrow time period. Nylon wool treatment of the cells eliminated the LPS response while SaGPIg and C' reduced it. Concanavalin A (con A) stimulated the cells under all test conditions and demonstrated a dose-time interrelationship in terms of maximum response. Pre-treatment of cells with SaGPIg and C' enhanced the response to con A while nylon wool fractionation diminished it somewhat. Only lymph node cells responded in vitro to oxidized ferredoxin (OFd). In serum-free media the OFd responses were maximal at 48 hours whereas in media containing FCS proliferative responses were supported for a prolonged period and appeared to be bimodal. Except for an early response with RPMI 1640 and ME, only media containing FCS supported stimulation in the mixed leucocyte culture.

摘要

在多种条件下,评估了豚鼠脾脏和淋巴结细胞在有丝分裂原(刀豆球蛋白A和脂多糖)、特异性抗原(氧化型铁氧还蛋白)以及同种异体细胞存在的情况下进行体外增殖反应的能力。在RPMI 1640、添加了巯基乙醇(ME)的RPMI 1640、添加了胎牛血清(FCS)的RPMI 1640以及含有ME和FCS的RPMI 1640中测量反应的时间和剂量依赖性。在用绵羊抗豚鼠免疫球蛋白(SaGPIg)和补体(C')处理细胞后,或在通过尼龙毛柱后,也测量了有丝分裂原反应。脂多糖(LPS)在所有培养基条件下的广泛浓度范围内均可刺激细胞,但作用时间较短。用尼龙毛处理细胞可消除LPS反应,而SaGPIg和C'则可使其降低。刀豆球蛋白A(Con A)在所有测试条件下均可刺激细胞,并在最大反应方面呈现剂量 - 时间相互关系。用SaGPIg和C'预处理细胞可增强对Con A的反应,而尼龙毛分级分离则使其有所减弱。只有淋巴结细胞在体外对氧化型铁氧还蛋白(OFd)有反应。在无血清培养基中,OFd反应在48小时时最大,而在含有FCS的培养基中,增殖反应可维持较长时间且似乎呈双峰。除了在RPMI 1640和ME存在下的早期反应外,只有含有FCS的培养基能支持混合白细胞培养中的刺激作用。

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Culture of mouse lymphoid cells in serum-free medium.在无血清培养基中培养小鼠淋巴细胞。
J Immunol Methods. 1972 Jan;1(2):199-202. doi: 10.1016/0022-1759(72)90046-4.

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