Assembling a Coculture System to Prepare Highly Pure Induced Pluripotent Stem Cell-Derived Neurons at Late Maturation Stages

被引:0
作者
Akter, Masuma [1 ]
Sepehrimanesh, Masood [1 ]
Xu, Wu [2 ]
Ding, Baojin [1 ]
机构
[1] Louisiana State Univ, Hlth Sci Ctr Shreveport, Dept Biochem & Mol Biol, Shreveport, LA 71130 USA
[2] Univ Louisiana Lafayette, Dept Chem, Lafayette, LA 70504 USA
基金
美国国家卫生研究院;
关键词
coculture; human induced pluripotent stem cells (hiPSCs); motor neurons (MNs); neurodevelopment; synaptogenesis; transcriptomics; MOTOR-NEURONS; DIFFERENTIATION; HIPPOCAMPAL; REQUIREMENT; EXPRESSION; GENERATION; OCCUPANCY; THERAPY;
D O I
10.1523/ENEURO.0165-24.2024
中图分类号
Q189 [神经科学];
学科分类号
071006 ;
摘要
Generation of human induced pluripotent stem cell (hiPSC)-derived motor neurons (MNs) offers an unprecedented approach to modeling movement disorders such as dystonia and amyotrophic lateral sclerosis. However, achieving survival poses a significant challenge when culturing induced MNs, especially when aiming to reach late maturation stages. Utilizing hiPSC-derived motor neurons and primary mouse astrocytes, we assembled two types of coculture systems: direct coculturing of neurons with astrocytes and indirect coculture using culture inserts that physically separate neurons and astrocytes. Both systems significantly enhance neuron survival. Compared with these two systems, no significant differences in neurodevelopment, maturation, and survival within 3 weeks, allowing to prepare neurons at maturation stages. Using the indirect coculture system, we obtained highly pure MNs at the late mature stage from hiPSCs. Transcriptomic studies of hiPSC-derived MNs showed a typical neurodevelopmental switch in gene expression from the early immature stage to late maturation stages. Mature genes associated with neurodevelopment and synaptogenesis are highly enriched in MNs at late stages, demonstrating that these neurons achieve maturation. This study introduces a novel tool for the preparation of highly pure hiPSC-derived neurons, enabling the determination of neurological disease pathogenesis in neurons at late disease onset stages through biochemical approaches, which typically necessitate highly pure neurons. This advancement is particularly significant in modeling age-related neurodegeneration.
引用
收藏
页数:14
相关论文
共 50 条
  • [21] Stimulation of GABA-Induced Ca2+ Influx Enhances Maturation of Human Induced Pluripotent Stem Cell-Derived Neurons
    Rushton, David J.
    Mattis, Virginia B.
    Svendsen, Clive N.
    Allen, Nicholas D.
    Kemp, Paul J.
    PLOS ONE, 2013, 8 (11):
  • [22] Epigenetic Priming of Human Pluripotent Stem Cell-Derived Cardiac Progenitor Cells Accelerates Cardiomyocyte Maturation
    Biermann, Mitch
    Cai, Wenxuan
    Lang, Di
    Hermsen, Jack
    Profio, Luke
    Zhou, Ying
    Czirok, Andras
    Isai, Dona G.
    Napiwocki, Brett N.
    Rodriguez, Adriana M.
    Brown, Matthew E.
    Woon, Marites T.
    Shao, Annie
    Han, Tianxiao
    Park, Donglim
    Hacker, Timothy A.
    Crone, Wendy C.
    Burlingham, William J.
    Glukhov, Alexey V.
    Ge, Ying
    Kamp, Timothy J.
    STEM CELLS, 2019, 37 (07) : 910 - 923
  • [23] Biowire: a platform for maturation of human pluripotent stem cell-derived cardiomyocytes
    Nunes, Sara S.
    Miklas, Jason W.
    Liu, Jie
    Aschar-Sobbi, Roozbeh
    Xiao, Yun
    Zhang, Boyang
    Jiang, Jiahua
    Masse, Stephane
    Gagliardi, Mark
    Hsieh, Anne
    Thavandiran, Nimalan
    Laflamme, Michael A.
    Nanthakumar, Kumaraswamy
    Gross, Gil J.
    Backx, Peter H.
    Keller, Gordon
    Radisic, Milica
    NATURE METHODS, 2013, 10 (08) : 781 - +
  • [24] Biowire platform for maturation of human pluripotent stem cell-derived cardiomyocytes
    Sun, Xuetao
    Nunes, Sara S.
    METHODS, 2016, 101 : 21 - 26
  • [25] A Toolbox to Characterize Human Induced Pluripotent Stem Cell-Derived Kidney Cell Types and Organoids
    Vanslambrouck, Jessica M.
    Wilson, Sean B.
    Tan, Ker Sin
    Soo, Joanne Y-C
    Scurr, Michelle
    Spijker, H. Siebe
    Starks, Lakshi T.
    Neilson, Amber
    Cui, Xiaoxia
    Jain, Sanjay
    Little, Melissa Helen
    Howden, Sara E.
    JOURNAL OF THE AMERICAN SOCIETY OF NEPHROLOGY, 2019, 30 (10): : 1811 - 1823
  • [26] Use of human induced pluripotent stem cell-derived neurons as a model for Cerebral Toxoplasmosis
    Tanaka, Naomi
    Ashour, Danah
    Dratz, Edward
    Halonen, Sandra
    MICROBES AND INFECTION, 2016, 18 (7-8) : 496 - 504
  • [27] Metabolomics Identifies Metabolic Markers of Maturation in Human Pluripotent Stem Cell-Derived Cardiomyocytes
    Bhute, Vijesh J.
    Bao, Xiaoping
    Dunn, Kaitlin K.
    Knutson, Kylie R.
    McCurry, Eric C.
    Jin, Gyuhyung
    Lee, Wei-Hua
    Lewis, Sarah
    Ikeda, Akihiro
    Palecek, Sean P.
    THERANOSTICS, 2017, 7 (07): : 2078 - 2091
  • [28] An Unbiased Proteomics Method to Assess the Maturation of Human Pluripotent Stem Cell-Derived Cardiomyocytes
    Cai, Wenxuan
    Zhang, Jianhua
    de Lange, Willem J.
    Gregorich, Zachery R.
    Karp, Hannah
    Farrell, Emily T.
    Mitchell, Stanford D.
    Tucholski, Trisha
    Lin, Ziqing
    Biermann, Mitch
    McIlwain, Sean J.
    Ralphe, J. Carter
    Kamp, Timothy J.
    Ge, Ying
    CIRCULATION RESEARCH, 2019, 125 (11) : 936 - 953
  • [29] Human Dendritic Cell-Derived Induced Pluripotent Stem Cell Lines Are Not Immunogenic
    Chhabra, Arvind
    Chen, I-Ping
    Batra, Deepika
    JOURNAL OF IMMUNOLOGY, 2017, 198 (05) : 1875 - 1886
  • [30] Role of somatic cell sources in the maturation degree of human induced pluripotent stem cell-derived cardiomyocytes
    Pianezzi, Enea
    Altomare, Claudia
    Bolis, Sara
    Balbi, Carolina
    Torre, Tiziano
    Rinaldi, Andrea
    Camici, Giovanni G.
    Barile, Lucio
    Vassalli, Giuseppe
    BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH, 2020, 1867 (03):