Derivation of Thymic Lymphoma T-cell Lines from Atm-/- and p53-/- Mice
Derivation of Thymic Lymphoma T-cell Lines from Atm-/- and p53-/- Mice
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DOI：
10.3791/2598
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发表时间：
2011-04-01
影响因子：
1.2
通讯作者：
Duhamel, Gerald
中科院分区：
文献类型：
--
作者：
Jinadasa, Rasika;Balmus, Gabriel;Duhamel, Gerald
建立细胞系是一种重要的研究工具，可以减少实验动物在研究中的使用。某些转基因小鼠品系，如Atm（-/-）和p53（-/-）在生命早期经常发生胸腺淋巴瘤（1,2），因此可以作为小鼠t细胞系衍生的可靠来源。在这里，我们提出了一种详细的方案，用于开发已建立的小鼠胸腺淋巴瘤t细胞系，而不需要像以前的方案那样添加白细胞介素（1,3）。肿瘤采集于3至6个月大的小鼠，根据观察弓背姿势、呼吸困难、梳理不良和易感品系消瘦，肿瘤最早出现可见迹象（1,4）。我们已经成功建立了多个t细胞系和Atm(-/-) [FVB/N-Atm(tm1Led)/J](2)和p53(-/-) [129/S6-Trp53(tm1Tyj)/J](5)小鼠近交系。我们进一步证明，超过90%的已建立的t细胞群表达CD3、CD4和CD8。与稳定建立的细胞系一致，使用本方案产生的t细胞已传代一年多。
Established cell lines are a critical research tool that can reduce the use of laboratory animals in research. Certain strains of genetically modified mice, such as Atm(-/-) and p53(-/-) consistently develop thymic lymphoma early in life (1,2), and thus, can serve as a reliable source for derivation of murine T-cell lines. Here we present a detailed protocol for the development of established murine thymic lymphoma T-cell lines without the need to add interleukins as described in previous protocols (1,3). Tumors were harvested from mice aged three to six months, at the earliest indication of visible tumors based on the observation of hunched posture, labored breathing, poor grooming and wasting in a susceptible strain (1,4). We have successfully established several T-cell lines using this protocol and inbred strains of Atm(-/-) [FVB/N-Atm(tm1Led)/J](2) and p53(-/-) [129/S6-Trp53(tm1Tyj)/J](5) mice. We further demonstrate that more than 90% of the established T-cell population expresses CD3, CD4 and CD8. Consistent with stably established cell lines, the T-cells generated by using the present protocol have been passaged for over a year.