Early-stage development of human induced pluripotent stem cell-derived neurons.
Recent advances in human induced pluripotent stem cells (hiPSCs) offer new possibilities for biomedical research and clinical applications. Differentiated neurons from hiPSCs are expected to be useful for developing novel methods of treatment for various neurological diseases. However, the detailed process of functional maturation of hiPSC-derived neurons (hiPS neurons) remains poorly understood. This study analyzes development of hiPS neurons, focusing specifically on early developmental stages through 48 hr after cell seeding; development was compared with that of primary cultured neurons derived from the rat hippocampus. At 5 hr after cell seeding, neurite formation occurs in a similar manner in both neuronal populations. However, very few neurons with axonal polarization were observed in the hiPS neurons even after 48 hr, indicating that hiPS neurons differentiate more slowly than rat neurons. We further investigated the elongation speed of axons and found that hiPS neuronal axons were slower. In addition, we characterized the growth cones. The localization patterns of skeletal proteins F-actin, microtubule, and drebrin were similar to those of rat neurons, and actin depolymerization by cytochalasin D induced similar changes in cytoskeletal distribution in the growth cones between hiPS neurons and rat neurons. These results indicate that, during the very early developmental stage, hiPS neurons develop comparably to rat hippocampal neurons with regard to axonal differentiation, but the growth of axons is slower.
Department of Neurobiology and Behavior, Gunma University Graduate School of Medicine, Maebashi, Gunma, Japan.
Yuki
Y
Ohara
Department of Neurobiology and Behavior, Gunma University Graduate School of Medicine, Maebashi, Gunma, Japan.
Noriko
N
Koganezawa
Department of Neurobiology and Behavior, Gunma University Graduate School of Medicine, Maebashi, Gunma, Japan.
Hiroyuki
H
Yamazaki
Department of Neurobiology and Behavior, Gunma University Graduate School of Medicine, Maebashi, Gunma, Japan.
Reiko T
RT
Roppongi
Division of Pharmacology, National Institute of Health Sciences, Tokyo, Japan.
Kaoru
K
Sato
Division of Pharmacology, National Institute of Health Sciences, Tokyo, Japan.
Yuko
Y
Sekino
Department of Neurobiology and Behavior, Gunma University Graduate School of Medicine, Maebashi, Gunma, Japan.
Tomoaki
T
Shirao
D000818:Animals; D001369:Axons / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D018920:Coculture Techniques; D015638:Cytochalasin D / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D004622:Embryo, Mammalian; D006624:Hippocampus / Q000166:cytology*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000502:physiology*; D007382:Intermediate Filaments / Q000378:metabolism; D018613:Microscopy, Confocal; D055495:Neurogenesis; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D009479:Neuropeptides / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013997:Time Factors; D014404:Tubulin / Q000378:metabolism
D009479:Neuropeptides; D014404:Tubulin; C053386:drebrins; D015638:Cytochalasin D
axonal development; cytoskeletal proteins; growth cones