Reviewer #3 (Public Review): Complete suspension culture of human induced pluripotent stem cells supplemented with suppressors of spontaneous differentiation Article Swipe

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Mami Matsuo‐Takasaki , Sho Kambayashi , Yasuko Hemmi , Tamami Wakabayashi , Tomoya Shimizu , Yuri An , Kazuhiro Takeuchi , Masato Ibuki , Yoshikazu Kawai , Masafumi Umekage , Tomoaki Kato , Michiya Noguchi , Koji Nakade , Yukio Nakamura , Tomoyuki Nakaishi , Masayoshi Tsukahara , Yohei Hayashi ·

YOU? · · 2023 · Open Access · · DOI: https://doi.org/10.7554/elife.89724.1.sa0

Human induced pluripotent stem cells (hiPSCs) are promising resources for producing various type of tissues in regenerative medicine; however, a scalable culture system that can precisely control the cell status for hiPSCs is not developed yet. Utilizing suspension culture without microcarriers or special materials allows for massive production, automation, cost effectiveness, and safety assurance in industrialized regenerative medicine. Here, we found that hiPSCs cultured in suspension conditions with continuous agitation without any microcarriers or extracellular matrix components were more prone to spontaneous differentiation than those cultured in conventional adherent conditions. Adding PKCβ and Wnt signaling pathway inhibitors in the suspension conditions suppressed the spontaneous differentiation of hiPSCs into ectoderm and mesendoderm, respectively. In these conditions, we successfully completed culture processes of hiPSCs including the generation of hiPSCs from peripheral blood mononuclear cells with the expansion of bulk population and single-cell sorted clones, long-term culture with robust self-renewal characteristics, single cell cloning, direct cryopreservation from suspension culture and their successful recovery, and efficient mass production of a clinical-grade hiPSC line. Our results demonstrate that precise control of hiPSC status in suspension culture conditions paves the way for their stable and automated clinical application.

Related Topics

Regenerative Medicine

Concepts

Microcarrier Regenerative medicine Induced pluripotent stem cell Cell biology Cell culture Human Induced Pluripotent Stem Cells Stem cell Biology Population Embryonic stem cell Biotechnology Chemistry Immunology Genetics Medicine Environmental health Gene

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Type: peer-review
Language: en
Landing Page: https://doi.org/10.7554/elife.89724.1.sa0
OA Status: gold
Related Works: 10
OpenAlex ID: https://openalex.org/W4390274464

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DOI: https://doi.org/10.7554/elife.89724.1.sa0

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Title: Reviewer #3 (Public Review): Complete suspension culture of human induced pluripotent stem cells supplemented with suppressors of spontaneous differentiation

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Type: peer-review

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Language: en

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Publication year: 2023

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Publication date: 2023-12-27

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Authors: Mami Matsuo‐Takasaki, Sho Kambayashi, Yasuko Hemmi, Tamami Wakabayashi, Tomoya Shimizu, Yuri An, Kazuhiro Takeuchi, Masato Ibuki, Yoshikazu Kawai, Masafumi Umekage, Tomoaki Kato, Michiya Noguchi, Koji Nakade, Yukio Nakamura, Tomoyuki Nakaishi, Masayoshi Tsukahara, Yohei Hayashi

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Landing page: https://doi.org/10.7554/elife.89724.1.sa0

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OA status: gold

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OA URL: https://doi.org/10.7554/elife.89724.1.sa0

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Concepts: Microcarrier, Regenerative medicine, Induced pluripotent stem cell, Cell biology, Cell culture, Human Induced Pluripotent Stem Cells, Stem cell, Biology, Population, Embryonic stem cell, Biotechnology, Chemistry, Immunology, Genetics, Medicine, Environmental health, Gene

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abstract_inverted_index.successful	158
abstract_inverted_index.suppressed	101
abstract_inverted_index.suspension	37, 65, 99, 154, 179
abstract_inverted_index.automation,	48
abstract_inverted_index.conditions,	114
abstract_inverted_index.conditions.	89
abstract_inverted_index.demonstrate	171
abstract_inverted_index.mononuclear	130
abstract_inverted_index.pluripotent	2
abstract_inverted_index.production,	47
abstract_inverted_index.single-cell	139
abstract_inverted_index.spontaneous	81, 103
abstract_inverted_index.application.	191
abstract_inverted_index.conventional	87
abstract_inverted_index.mesendoderm,	110
abstract_inverted_index.regenerative	16, 56
abstract_inverted_index.self-renewal	146
abstract_inverted_index.successfully	116
abstract_inverted_index.extracellular	74
abstract_inverted_index.microcarriers	40, 72
abstract_inverted_index.respectively.	111
abstract_inverted_index.clinical-grade	166
abstract_inverted_index.effectiveness,	50
abstract_inverted_index.industrialized	55
abstract_inverted_index.differentiation	82, 104
abstract_inverted_index.characteristics,	147
abstract_inverted_index.cryopreservation	152
cited_by_percentile_year
countries_distinct_count	0
institutions_distinct_count	17
sustainable_development_goals[0].id	https://metadata.un.org/sdg/9
sustainable_development_goals[0].score	0.4099999964237213
sustainable_development_goals[0].display_name	Industry, innovation and infrastructure
citation_normalized_percentile