张理超, 张捷, 付必莽, 赵越, 唐波, 陈贵元, 王维, 李梅章. 树鼩淋巴细胞的快速分离和分析方法的初步建立[J]. 云南大学学报(自然科学版), 2013, 35(S2): 359. doi: 10.7540/j.ynu.20130583
引用本文: 张理超, 张捷, 付必莽, 赵越, 唐波, 陈贵元, 王维, 李梅章. 树鼩淋巴细胞的快速分离和分析方法的初步建立[J]. 云南大学学报(自然科学版), 2013, 35(S2): 359. doi: 10.7540/j.ynu.20130583
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Establishment of quick isolation and analysisof tree shrew’s lymphocytes[J]. Journal of Yunnan University: Natural Sciences Edition, 2013, 35(S2): 359. DOI: 10.7540/j.ynu.20130583
Citation: Establishment of quick isolation and analysisof tree shrew’s lymphocytes[J]. Journal of Yunnan University: Natural Sciences Edition, 2013, 35(S2): 359. DOI: 10.7540/j.ynu.20130583
shu

树鼩淋巴细胞的快速分离和分析方法的初步建立

Establishment of quick isolation and analysisof tree shrew’s lymphocytes

    摘要
  • 摘要: 本研究旨在建立树鼩淋巴细胞分离和培养的方法.根据组织来源，本实验分为3个实验组，第1组，采用树鼩活体改良心脏采血法并联合密度梯度离心法分离树鼩淋巴细胞；第2组，CO2处死树鼩，采用逆向解剖法游离其胫骨股骨制备骨髓细胞；第3组，直接获取脾脏细胞.上述3种方法收集过的淋巴细胞，经过12h培养后，先用台盼蓝染色检测细胞的存活率，再应用流式细胞技术分析淋巴细胞的活力.结果显示3种方法所获取的树鼩淋巴细胞所占比例各异，具有组织特异性.0.5mL的树鼩外周血可获得淋巴细胞大约为6.25×104 个，占细胞总数的50%；双侧胫骨股骨可获得淋巴细胞大约为1.96×105 个，占细胞总数的6.31%；树鼩脾脏可获得淋巴细胞大约为1.59×106 个，占细胞总数的23.7%.各方法所获得细胞活力良好，且在流式细胞技术检测中可见明显分群.本研究初步建立快速分离和培养树鼩淋巴细胞的方法，为树鼩后续的淋巴细胞的免疫研究打下坚实的基础.

     

    Abstract: To study the lymphocytes from tree shrew,we established and developed three quick protocols to isolate and analyze the cells from peripheral blood,bone and spleen tissues.Peripheral blood was collected by live heart puncture and lymphocytes were further isolated by mouse Ficoll-Hypaque.Lymphocytes were also separated from both bone marrow and spleen.About 6.25×104,1.96×105 and 1.59×106 lymphocytes were obtained from blood,bone marrow and spleen respectively.Results from flow cytometry analysis further confirmed the successful isolation of tree shrew’s lymphocytes.

     

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