上海脑库脑组织样本的质量控制分析  

Quality control analysis of brain tissue samples from Shanghai Brain Bank

作　　者：李凤娇 咸伟伟 卜硕蕾 曹静丽 吴劲松 李文生 尤琳雅 LI Feng-jiao;XIAN Wei-wei;BU Shuo-lei;CAO Jing-li;WU Jin-song;LI Wen-sheng;YOU Lin-ya(Department of Human Anatomy and Histoembryology,School of Basic Medical Sciences,Fudan University,Shanghai 200032,China;Institutes of Biomedical Sciences,Fudan University,Shanghai 200032,China;Department of Neurosurgery,Huashan Hospital,Fudan University,Shanghai 200040,China;Shanghai Key Laboratory of Medical Imaging Computing and Computer Assisted Intervention,Shanghai 200032,China)

机构地区：[1]复旦大学基础医学院人体解剖学与组织学胚胎学系,上海200032 [2]复旦大学生物医学研究院,上海200032 [3]复旦大学附属华山医院神经外科,上海200040 [4]上海市医学图像处理与计算机辅助手术重点实验室,上海200032

出　　处：《解剖学报》2021年第2期168-174,共7页Acta Anatomica Sinica

基　　金：国家自然科学基金(81771228);复旦大学“卓越2025”卓学优秀计划(JIH1829604);上海脑库技术体系的初步建设项目(16JC1420103)。

摘　　要：目的通过对脑脊液pH值、脑组织RNA完整值(RIN)、转录组和蛋白质组检测以及形态学观察,评估上海脑库收集的脑组织用于后续生物学研究的可行性。方法使用pH试纸检测新鲜脑脊液pH值;使用安捷伦RNA 6000 Nano芯片和2100生物分析仪对低温冻存的脑组织RNA进行完整性检测;使用BGIseq-500测序仪对低温冻存的颞上回(STG)或尾状核(CN)进行转录组测序;使用Q Exactive HF质谱仪对低温冻存脑组织进行蛋白质组分析;使用HE染色观察STG的形态学。结果捐献脑组织样本的脑脊液pH值在6.5左右;65%以上的低温冻存脑组织的RIN>6,RNA质量较好;转录组测序过滤后clean reads比值>20或30的碱基数占总碱基数比例基本达到80%以上,表明文库质量较高;质谱分析得到超过4000个蛋白质组,3万多个肽段,说明蛋白数据质量较好;脑组织形态学较为正常,能清晰观察到神经元形态。结论上海脑库脑组织样本的RNA和蛋白质质量能用于开展分子生物学相关研究,并可用于形态学观察。Objective To evaluate the feasibility of using brain tissues from Shanghai Brain Bank for the applications on biological research through the analysis of pH value of cerebrospinal fluids,RNA integrity number(RIN),transcriptome,proteome and morphology of brain tissues.Methods The pH value of fresh cerebrospinal fluid was detected by pH test paper;the RNA integrity of cryopreserved brain tissues was examined by Agilent RNA 6000 Nano chip and Agilent 2100 bioanalyzer;the transcriptome sequencing of superior temporal gyrus or caudate was performed using BGIseq-500 sequencer;the proteome of cryopreserved brain tissues was analyzed by Q Exactive HF mass spectrometer;at last,the morphology of the superior temporal gyrus was observed by HE staining.Results The pH value of the cerebrospinal fluid on average was about 6.5.The RIN values of more than 65%of brain tissues were more than 6,indicating good RNA quality.The clean reads ratio after transcriptome sequencing filtering was basically above 80%,indicating that the quality of sequencing library was high.The mass spectrometry analysis of frozen brain samples yielded more than 4000 protein groups and 30000 peptides,indicating high quality of proteomic data.The morphology of brain tissues was relatively normal,with clearly visible neurons.Conclusion The quality of RNA and protein of brain tissues from Shanghai Brain Bank meets the basic needs for molecular and biological research,and the fixed brain samples can be used for morphological observations.

关 键 词：脑脊液酸碱度 脑组织 核糖核酸 蛋白质 HE染色 人 

分 类 号：R322.81[医药卫生—人体解剖和组织胚胎学]