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.
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页数:14
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