Controlled Growth and the Maintenance of Human Pluripotent Stem Cells by Cultivation with Defined Medium on Extracellular Matrix-Coated Micropatterned Dishes

Takenaka, Chiemi; Miyajima, Hiroshi; Yoda, Yusuke; Imazato, Hideo; Yamamoto, Takako; Gomi, S.; Ohshima, Yasuhiro; Kagawa, Kenichi; Sasaki, Tomohiro; Kawamata, Shin

doi:10.1371/journal.pone.0129855

Cited by 13 publications

(11 citation statements)

References 18 publications

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“…Human ESC lines KhES-1 15 (Riken BRC) and H9 16 (WiCell), and the human iPSC lines PFX#9 17 , 201B7 18 (Riken BRC) and SHh#2 17 were cultured with hPSCs culture media on mitomycin C-treated SNL76/7 cells (SIM strain embryonic fibroblast, European Collection of Cell Culture) or with chemical defined medium Essential 8 19 (Es8, Thermo Fisher Scientific), serum-free medium Stem-Partner 20 (S-P, Kyokuto Pharma), chemical defined medium Stem-Partner ACF (S-P ACF, Kyokuto Pharma) or xeno-free Repro FF2 medium (ReproCELL) on recombinant human Vitronectin-N (Thermo Fisher Scientific)-coated dishes. Cells were passaged in clumps using Gentle Cell Dissociation Reagent (GCDR; Stem Cell Technologies) and split at a ratio of 1:3 for iPSCs or 1:3.5 for ESCs.…”

Section: Discussionmentioning

confidence: 99%

Differentiation potential of Pluripotent Stem Cells correlates to the level of CHD7

Yamamoto

Takenaka

Yoda

et al. 2018

Sci Rep

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Embryonic Stem Cells (ESC) possesses two distinct features; self-renewal and the potential to differentiate. Here we show the differentiation potential and growth rate of ESC correlates positively with the expression level of the gene encoding chromodomain helicase DNA binding protein 7 (CHD7). When ESCs are maintained in feeder-free conditions and single cell seeding, ESC KhES-1 having 4520 copies or more of CHD7 in 5 ng total RNA show differentiation potential, but this is lost when the CHD7 copy number is reduced in KhES-1 to less than 696 by alternative culture conditions. Introduction of siCHD7 reduced differentiation potential and growth rate of KhES-1. Interestingly, KhES-1 underwent spontaneous differentiation when mCHD7 was introduced and we could not obtain CHD7-overexpressing ESC in culture. These data suggest that CHD7 drives differentiation, and there is a lower limit for CHD7 to initiate differentiation and an upper limit for CHD7 if maintained in undifferentiated state, and such upper limit varies depending on culture condition. As CHD7 drives cell growth, ESC with the highest permissible CHD7 level in the given culture become dominant in a couple of passages. Thus, we can select differentiation resistance-free cell clones by optimizing the culture system using CHD7 as an index.

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Section: Discussionmentioning

confidence: 99%

Differentiation potential of Pluripotent Stem Cells correlates to the level of CHD7

Yamamoto

Takenaka

Yoda

et al. 2018

Sci Rep

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“…Synthetic culture surfaces such as recombinant human vitronectin-N-coated dishes or biomaterial coating [326] Xeno-free nutrition supplements such as ThermoFisher Scientific N2 (A1370701) and B27 (A3353501)…”

Section: Risksmentioning

confidence: 99%

Advances in Pluripotent Stem Cells: History, Mechanisms, Technologies, and Applications

Liu

David

Trawczynski

et al. 2019

Stem Cell Rev and Rep

387

209

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Over the past 20 years, and particularly in the last decade, significant developmental milestones have driven basic, translational, and clinical advances in the field of stem cell and regenerative medicine. In this article, we provide a systemic overview of the major recent discoveries in this exciting and rapidly developing field. We begin by discussing experimental advances in the generation and differentiation of pluripotent stem cells (PSCs), next moving to the maintenance of stem cells in different culture types, and finishing with a discussion of three-dimensional (3D) cell technology and future stem cell applications. Specifically, we highlight the following crucial domains: 1) sources of pluripotent cells; 2) next-generation in vivo direct reprogramming technology; 3) cell types derived from PSCs and the influence of genetic memory; 4) induction of pluripotency with genomic modifications; 5) construction of vectors with reprogramming factor combinations; 6) enhancing pluripotency with small molecules and genetic signaling pathways; 7) induction of cell reprogramming by RNA signaling; 8) induction and enhancement of pluripotency with chemicals; 9) maintenance of pluripotency and genomic stability in induced pluripotent stem cells (iPSCs); 10) feeder-free and xenon-free culture environments; 11) biomaterial applications in stem cell biology; 12) three-dimensional (3D) cell technology; 13) 3D bioprinting; 14) downstream stem cell applications; and 15) current ethical issues in stem cell and regenerative medicine. This review, encompassing the fundamental concepts of regenerative medicine, is intended to provide a comprehensive portrait of important progress in stem cell research and development. Innovative technologies and real-world applications are emphasized for readers interested in the exciting, promising, and challenging field of stem cells and those seeking guidance in planning future research direction.

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“…17,19,20 Different topographical cues have been tested already to increase hiPSC-CM elongation, like micropatterned substrates, 21 substrate stiffness, 17 and microgrids. 19,20 However, the ideal spacing and depth combination of a microtopography to mature hiPSC-CM has not been examined yet.…”

Section: Introductionmentioning

confidence: 99%

Enhanced Human-Induced Pluripotent Stem Cell Derived Cardiomyocyte Maturation Using a Dual Microgradient Substrate

Huethorst

Hortigon

Zamora

et al. 2016

ACS Biomater. Sci. Eng.

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Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) raise many possibilities for cardiac research but they exhibit an immature phenotype, which influences experimental outcomes. The aim of our research is to investigate the effects of a topographical gradient substrate on the morphology and function of commercially available hiPSC-CM. The lateral dimensions the microgrooves on the substrate varied from 8 to 100 μm space between the 8 μm grooves on one axis and from ∼5 nm to ∼1 μm in depth on the other axis. Cells were seeded homogeneously across the substrate and according to the manufacturers protocols. At days 4 and 10, measures of eccentricity, elongation, orientation, sarcomere length (SL), and contractility of the hiPSC-CM were taken. Only the deepest and widest region (8–30 μm wide and 0.85–1 μm deep) showed a significantly higher percentage of hiPSC-CM with an increased eccentricity (31.3 ± 6.4%), elongation (10.4 ± 4.3%), and orientation (<10°) (32.1 ± 2.7%) when compared with the control (flat substrate) (15.8 ± 5.0%, 3.4 ± 2.7%, and 10.6 ± 1.1%, respectively). Additionally, during stimulus-induced contraction, the relaxation phase of the twitch was prolonged (400 ms) compared to nonelongated cells (200 ms). These findings support the potential use of dual microgradient substrates to investigate substrate topographies that stimulate migration and/or maturation of hiPSC-CM.

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Controlled Growth and the Maintenance of Human Pluripotent Stem Cells by Cultivation with Defined Medium on Extracellular Matrix-Coated Micropatterned Dishes

Cited by 13 publications

References 18 publications

Differentiation potential of Pluripotent Stem Cells correlates to the level of CHD7

Differentiation potential of Pluripotent Stem Cells correlates to the level of CHD7

Advances in Pluripotent Stem Cells: History, Mechanisms, Technologies, and Applications

Enhanced Human-Induced Pluripotent Stem Cell Derived Cardiomyocyte Maturation Using a Dual Microgradient Substrate

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