首页> 外文期刊>Egyptian Journal of Medical Human Genetics >A modified protocol for highly efficient EBV-mediated immortalization of human B lymphocytes from small volumes of peripheral blood serum
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A modified protocol for highly efficient EBV-mediated immortalization of human B lymphocytes from small volumes of peripheral blood serum

机译:改良的协议,可从少量外周血中高效EBV介导的人类B淋巴细胞永生化

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Background Many human molecular and genetic studies require the use of a renewable biological material. Although primary fibroblast cell lines can be used for this purpose, there are disadvantages associated with human biopsies including the limited number of cell divisions. Peripheral blood has the advantage of being easier to obtain but also the drawback that blood cells produce only short-term cultures. Epstein-Barr virus is capable of transforming human B lymphocytes into indefinitely proliferating cells that can be maintained in tissue culture. Here, we report a convenient method of B-lymphocyte immortalization using small volumes of freshly collected human blood serum saturated with nucleated blood cells. Aim of the study The aim of the present study is modification and improvement of the protocol for highly efficient immortalization of human B lymphocytes from small volume of blood samples. Material and methods Cell line B95-8 was used as Epstein-Barr virus source for viral stock preparation. Immortalizing medium contains RPMI-1640, viral stock and additives. No feeder layer was used. Results As result we present high efficient method for B lymphocytes immortalization with start blood volume less than 5?ml. Conclusion The method is applicable for immortalization of B lymphocytes from small blood samples and is essential for studies involving children or patients when large blood volume sampling is impossible.
机译:背景技术许多人类分子和遗传研究要求使用可再生生物材料。尽管原代成纤维细胞系可用于此目的,但与人类活组织检查有关的缺点包括有限的细胞分裂数目。外周血的优点是易于获取,但缺点是血细胞仅产生短期培养物。爱泼斯坦-巴尔病毒能够将人B淋巴细胞转化为无限增殖的细胞,可以在组织培养物中维持。在这里,我们报告了使用少量新鲜收集的,有核血细胞饱和的人血清的B淋巴细胞永生化的便捷方法。研究目的本研究的目的是改进和改进从少量血样中高效永生化人类B淋巴细胞的方案。材料和方法细胞系B95-8被用作制备病毒原种的爱泼斯坦-巴尔病毒源。永生培养基包含RPMI-1640,病毒原液和添加剂。没有使用饲养层。结果结果,我们提出了一种有效的B淋巴细胞永生化方法,起始血容量小于5?ml。结论该方法适用于永生化小血样中的B淋巴细胞,对于无法进行大血样取样的儿童或患者的研究至关重要。

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