CFSE标记神经祖细胞方法的建立与验证  

Establishment and validation of a method labelling neural progenitor cells with CFSE

作　　者：周佳锋 马孟杰 彭小忠[1,2] 舒鹏程 ZHOU Jia-feng;MA Meng-jie;PENG Xiao-zhong;SHU Peng-cheng(State Key Laboratory of Medical Molecular Biology, Department of Molecular Biology and Biochemistry,Medical Primate Center, Neuroscience Center, Institute of Basic Medical Sciences CAMS, School of Basic Medicine PUMC,Beijing 100005;Institute of Medical Biology CAMS, Kunming 650118, China)

机构地区：[1]中国医学科学院基础医学研究所,北京协和医学院基础学院,生物化学与分子生物学系,医学分子生物学国家重点实验室医学灵长类研究中心神经科学中心,北京100005 [2]中国医学科学院医学生物学研究所,云南昆明650118

出　　处：《基础医学与临床》2021年第5期648-652,共5页Basic and Clinical Medicine

基　　金：国家自然科学基金(31970772)。

摘　　要：目的建立并验证羧基荧光素二醋酸盐琥珀酰亚胺脂(CFSE)荧光探针标记神经祖细胞(NPCs)的方法。方法在胚胎期小鼠侧脑室注射CFSE以标记室周区(VZ)细胞,在注射后的1 h和3 h分别收集组织,通过免疫荧光方法检测其特异性。在注射后的1 h分离出背侧新皮质,通过流式细胞荧光分选术(FACS)分选强阳性细胞,并通过干细胞标志物、神经元标志物等验证。结果CFSE能够瞬时标记VZ区细胞,标记窗在3~6 h。CFSE进行长时程标记会出现衰减。利用CFSE分选所得细胞70%为神经祖细胞,少量为神经元等。结论CFSE能够用于瞬时及长时程在体标记。CFSE还可用于分选神经祖细胞,但尚需调整分选策略以提高特异性(FACS)。Objective Establishing and verifying a method for labeling neural progenitor cells(NPCs)with carboxyl fluorescein diacetate succinimidyl ester(CFSE)fluorescent probe.Methods CFSE was injected into the lateral ventricle of embryonic mice to label ventricular zone(VZ)cells.The tissue was collected at 1 and 3 hours after injection to detect its specificity by immunofluorescence.Dorsal neocortex was separated at 1 hour after injection,the strong positive cells were sorted by fluorescence activated cell sorting(FACS)and the cell identity was verified by stem cell markers and neuronal markers.Results CFSE instantaneously labelled(pulse labeling)cells in the ventricular zone,and the labeling window was between 3 and 6 hours.Signal of CFSE when performing long-term labeling would be attenuated.Seventy percent of the cells sorted by CFSE were neural precursor cells,and a small portion of which were neurons.Conclusions CFSE can be used for pulse and long-term labeling in vivo.Also,CFSE can be applied for sorting NPCs,while the current sorting strategy needs to be adjusted to improve specificity.

关 键 词：羧基荧光素二醋酸盐琥珀酰亚胺脂(CFSE) 神经祖细胞 流式细胞荧光分选术(FACS) 

分 类 号：Q71[生物学—分子生物学]