In vitro models of neuronal aging and gene manipulation in human neurons (hNeurons) are valuable tools for investigating human brain aging and diseases. Here, we present a protocol for applying human embryonic stem cell (hESC)derived neurons to model aging and the further application of small interfering RNA (siRNA)-mediated gene silencing for functional investigations. We describe steps for neuronal differentiation and culture, siRNA transfection, and technical considerations to ensure reproducibility. Our protocol enables investigations of the molecular mechanism underlying neuronal aging and facilitates drug evaluation. For complete details on the use and execution of this protocol, please refer to Zhang et al.1
基金：
National Key Research and Development Program of China [2020YFA0804 000, 2022YFA1103700, 2020YFA0804000]; National Natural Science Foundation of China [82330044, 32341001, 32121001, 82192863, 82361148130, 8231101626, 82488301, 82125011, 92168201, 82361148131]; Program of the Beijing Natural Science Foundation [JQ24044, Z240018, Z230011]; Shenzhen Medical Research Fund [C2406001]; CAS Project for Young Scientists in Basic Research [YSBR-076, YSBR-012]; Informatization Plan of Chinese Academy of Sciences [CAS-WX2022SDC-XK14]; New Cornerstone Science Foundation [2021-1045]; Beijing Municipal Public Welfare Development and Reform Pilot Project for Medical Research Institutes [JYY2023-13]; CAS Youth Interdisciplinary Team, Space Medical Experiment Project of CMSP [HYZHXMH01012]; Key Laboratory of Alzheimer's Disease of Zhejiang Province [ZJAD-2024001]; Initiative Scientific Research Program, Institute of Zoology, CAS [2023IOZ0202]; Excellent Young Talents Program of Capital Medical University [12300927]; Excellent Young Talents Training Program for the Construction of Beijing Municipal University Teacher Team [BPHR202203105]
第一作者机构：[1]Capital Med Univ, Beijing Anzhen Hosp, Beijing Inst Heart Lung & Blood Vessel Dis, Beijing 100029, Peoples R China
共同第一作者：
通讯作者：
通讯机构：[1]Capital Med Univ, Beijing Anzhen Hosp, Beijing Inst Heart Lung & Blood Vessel Dis, Beijing 100029, Peoples R China[2]Chinese Acad Sci, Inst Zool, Key Lab Organ Regenerat & Reconstruct, State Key Lab Membrane Biol,State Key Lab Stem Cel, Beijing 100101, Peoples R China[3]Beijing Inst Stem Cell & Regenerat Med, Beijing 100101, Peoples R China[4]Capital Med Univ, Xuanwu Hosp, Natl Clin Res Ctr Geriatr Disorders, Aging Translat Med Ctr, Beijing 100053, Peoples R China[5]Univ Chinese Acad Sci, Beijing 100049, Peoples R China[6]China Natl Ctr Bioinformat, Beijing, Peoples R China[7]Chinese Acad Sci, Beijing Inst Genom, Beijing, Peoples R China[8]Aging Biomarker Consortium ABC, Beijing 100101, Peoples R China
推荐引用方式(GB/T 7714)：
Zhang Hui,Sun Shuhui,Belmonte Juan Carlos Izpisua,et al.Protocols for the application of human embryonic stem cell-derived neurons for aging modeling and gene manipulation[J].STAR PROTOCOLS.2025,6(1):doi:10.1016/j.xpro.2025.103633.
APA：
Zhang, Hui,Sun, Shuhui,Belmonte, Juan Carlos Izpisua,Liu, Guang-Hui,Wang, Si...&Qu, Jing.(2025).Protocols for the application of human embryonic stem cell-derived neurons for aging modeling and gene manipulation.STAR PROTOCOLS,6,(1)
MLA：
Zhang, Hui,et al."Protocols for the application of human embryonic stem cell-derived neurons for aging modeling and gene manipulation".STAR PROTOCOLS 6..1(2025)
