scholarly journals Fluorescent indicators for continuous and lineage‐specific reporting of cell‐cycle phases in human pluripotent stem cells

2020 ◽  
Vol 117 (7) ◽  
pp. 2177-2186
Author(s):  
Yun Chang ◽  
Peter B. Hellwarth ◽  
Lauren N. Randolph ◽  
Yufei Sun ◽  
Yuxian Xing ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Human Pluripotent Stem Cells ◽  
Fluorescent Indicators ◽  
Cell Cycle Phases

scholarly journals A Src inhibitor regulates the cell cycle of human pluripotent stem cells and improves directed differentiation

2015 ◽  
Vol 210 (7) ◽  
pp. 1257-1268 ◽  
Author(s):  
Sundari Chetty ◽  
Elise N. Engquist ◽  
Elie Mehanna ◽  
Kathy O. Lui ◽  
Alexander M. Tsankov ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Human Pluripotent Stem Cells ◽  
Directed Differentiation ◽  
Differentiation Potential ◽  
Germ Layers ◽  
Positive Effects ◽  
Src Inhibitor ◽  
Expression Of Genes

Driving human pluripotent stem cells (hPSCs) into specific lineages is an inefficient and challenging process. We show that a potent Src inhibitor, PP1, regulates expression of genes involved in the G1 to S phase transition of the cell cycle, activates proteins in the retinoblastoma family, and subsequently increases the differentiation propensities of hPSCs into all three germ layers. We further demonstrate that genetic suppression of Src regulates the activity of the retinoblastoma protein and enhances the differentiation potential of hPSCs across all germ layers. These positive effects extend beyond the initial germ layer specification and enable efficient differentiation at subsequent stages of differentiation.

scholarly journals A Src inhibitor regulates the cell cycle of human pluripotent stem cells and improves directed differentiation

2015 ◽  
Vol 212 (11) ◽  
pp. 21211OIA91 ◽  
Author(s):  
Sundari Chetty ◽  
Elise Engquist ◽  
Elie Mehanna ◽  
Kathy Lui ◽  
Alexander Tsankov ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Human Pluripotent Stem Cells ◽  
Directed Differentiation ◽  
Src Inhibitor

scholarly journals Cell cycle dynamics of human pluripotent stem cells primed for differentiation

2019 ◽  
Author(s):  
Anna Shcherbina ◽  
Jingling Li ◽  
Cyndhavi Narayanan ◽  
William Greenleaf ◽  
Anshul Kundaje ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Expression Patterns ◽  
Human Pluripotent Stem Cells ◽  
Specific Manner ◽  
A Cell ◽  
Cell Cycle Dynamics ◽  
Differentiation Gene ◽  
Cycle Indicator

Understanding the molecular properties of the cell cycle of human pluripotent stem cells (hPSCs) is critical for effectively promoting differentiation. Here, we use the Fluorescence Ubiquitin Cell Cycle Indicator (FUCCI) system adapted into hPSCs and perform RNA-sequencing on cell cycle sorted hPSCs primed and unprimed for differentiation. Gene expression patterns of signaling factors and developmental regulators change in a cell cycle-specific manner in cells primed for differentiation without altering genes associated with pluripotency. Furthermore, we identify an important role for PI3K signaling in regulating the early transitory states of hPSCs towards differentiation.

scholarly journals Establish deeper understanding on the osteogenic differentiation of monolayer cultured human pluripotent stem cells using novel and detail analyses

2020 ◽  
Author(s):  
Ping Zhou ◽  
Jia-Min Shi ◽  
Jing-E Song ◽  
Yu Han ◽  
Hong-Jiao Li ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Osteogenic Differentiation ◽  
Pluripotent Stem Cells ◽  
Telomerase Activity ◽  
Human Pluripotent Stem Cells ◽  
Cell Counting ◽  
Integrated Analysis ◽  
Differentiation Process ◽  
Cell Type

Abstract Background: Derivation of the osteoblast-like cell from human pluripotent stem cell becomes a hot topic in bone tissue engineering. Although many improvements have been achieved in this field, low induction efficiency because of the non-directed differentiation process hampers their application in bone regeneration. We think lack of detailed understanding on the osteogenic differentiation process should be the main reason.Methods: Monolayer cultured human embryonic stem cells and human induced pluripotent stem cells were inducted in traditional serum-containing osteogenic medium for 35 days. Except for traditional assays such as cell viability detection, reverse transcription-polymerase chain reaction, immunofluorescence, and alizarin red staining, we also applied cell counting, cell telomerase activity, cell cycle and quantitative expression of runt-related transcription factor 2 as essential indicators to analyze the cell type changes during the differentiation process. Results: The population of differentiated cells are quite heterogenous throughout 35 days of induction. Then, cell telomerase activity and cell cycle analyses have value in evaluating the cell type changes and tumorigenicity of obtained cells. Moreover, nuclear staining should be a recommended method to evaluate the cell number, because, it is still a great challenge to dissociate cells with varying differentiation times into single cells with high survival rate. Finally, a dynamic map was made to integrated analysis of these results, and the cell types at defined stages of osteogenic differentiation of human pluripotent stem cells was concluded. Conclusions: This study lay foundation to improve the in vitro osteogenic differentiation efficiency of human pluripotent stem cells by supplementing functional compounds at each stage according to a time-frame, then establish a step-wised induction system in the future.

RELB Alters Proliferation of Human Pluripotent Stem Cells via IMP3- and LIN28-Mediated Modulation of the Expression of IGF2 and Other Cell-Cycle Regulators

2015 ◽  
Vol 24 (16) ◽  
pp. 1888-1900 ◽  
Author(s):  
Nilay Yogeshkumar Thakar ◽  
Dmitry Alexander Ovchinnikov ◽  
Marcus Lachlan Hastie ◽  
Jeffrey Gorman ◽  
Ernst Jurgen Wolvetang
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Human Pluripotent Stem Cells ◽  
Cell Cycle Regulators

scholarly journals TDG regulates cell cycle progression in human neural progenitors

F1000Research ◽  
2018 ◽  
Vol 7 ◽  
pp. 497
Author(s):  
Igal Germanguz ◽  
Jenny C. Park ◽  
Jessica Cinkornpumin ◽  
Aryeh Solomon ◽  
Minori Ohashi ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Dna Methylation ◽  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Methylation Status ◽  
Human Pluripotent Stem Cells ◽  
Dna Demethylation ◽  
Neural Progenitors ◽  
Specific Cell ◽  
Active Demethylation

Background: As cells divide, they must both replicate their DNA and generate a new set of histone proteins.  The newly synthesized daughter strands and histones are unmodified, and must therefore be covalently modified to allow for transmission of important epigenetic marks to daughter cells.  Human pluripotent stem cells (hPSCs) display a unique cell cycle profile, and control of the cell cycle is known to be critical for their proper differentiation and survival.  A major unresolved question is how hPSCs regulate their DNA methylation status through the cell cycle, namely how passive and active demethylation work to maintain a stable genome. Thymine-DNA glycosylase (TDG), an embryonic essential gene, has been recently implicated as a major enzyme involved in demethylation. Methods: We use human pluripotent stem cells and their derivatives to investigate the role of TDG in differentiation and proliferation.  To perform loss of function of TDG, RNA Interference was used.  To study the cell cyle, we engineered human pluripotent stem cells to express the FUCCI tool which marks cells at various stages of the cell cycle with distinct patterns of fluorescent proteins.  We also used cell cycle profiling by FACS, and DNA methylation analysis to probe a connection between DNA demethylation and cell cycle. Results: Here we present data showing that TDG regulates cell cycle dynamics in human neural progenitors (NPCs) derived from hPSCs, leading to changes in  cell cycle related gene expression and neural differentiation capacity.  These data show that loss of TDG function can block differentiation by driving proliferation of neural progenitors.  We also identify specific cell cycle related genes whose expression changes upon loss of TDG expression. Conclusions: These observations suggest that TDG and active demethylation play an important role in hPSC cell cycle regulation and differentiation.

scholarly journals MicroRNA miR-302 Inhibits the Tumorigenecity of Human Pluripotent Stem Cells by Coordinate Suppression of the CDK2 and CDK4/6 Cell Cycle Pathways

2010 ◽  
Vol 70 (22) ◽  
pp. 9473-9482 ◽  
Author(s):  
Shi-Lung Lin ◽  
Donald C. Chang ◽  
Shao-Yao Ying ◽  
Davey Leu ◽  
David T.S. Wu
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Human Pluripotent Stem Cells

scholarly journals Cell cycle regulators control mesoderm specification in human pluripotent stem cells

2019 ◽  
Author(s):  
Loukia Yiangou ◽  
Rodrigo A. Grandy ◽  
Sanjay Sinha ◽  
Ludovic Vallier
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Human Pluripotent Stem Cells ◽  
Bmp Signaling ◽  
Dependent Manner ◽  
Cell Cycle Regulators ◽  
Lateral Plate ◽  
Mesoderm Specification ◽  
Mesoderm Formation

ABSTRACTMesoderm is one of the three germ layers produced during gastrulation from which muscle, bones, kidneys and the cardiovascular system originate. Understanding the mechanisms controlling mesoderm specification could be essential for a diversity of applications, including the development of regenerative medicine therapies against diseases affecting these tissues. Here, we use human pluripotent stem cells (hPSCs) to investigate the role of cell cycle in mesoderm formation. For that, proteins controlling G1 and G2/M cell cycle phases were inhibited during differentiation of hPSCs into lateral plate, cardiac and presomitic mesoderm using small molecules or by conditional knock down. These loss of function experiments revealed that CDKs and pRb phosphorylation are necessary for efficient mesoderm formation in a context-dependent manner. Further investigations showed that inhibition of the G2/M regulator CDK1 decreases BMP signaling activity specifically during lateral plate mesoderm formation while reducing FGF/ERK1/2 activity in all mesoderm subtypes. Taken together, our findings reveal that cell cycle regulators direct mesoderm formation by controlling the activity of key developmental pathways.

miR-302 family, miR-145 and miR-296 temporal expression profile along the cell cycle of human pluripotent stem cells

2021 ◽  
Vol 40 ◽  
pp. 119168
Author(s):  
María Soledad Rodríguez-Varela ◽  
Sofía Mucci ◽  
Guillermo Agustín Videla-Richardson ◽  
Luciana Isaja ◽  
Gustavo Emilio Sevlever ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Expression Profile ◽  
Pluripotent Stem Cells ◽  
Human Pluripotent Stem Cells ◽  
Temporal Expression ◽  
Temporal Expression Profile

scholarly journals Enrichment of G2/M cell cycle phase in human pluripotent stem cells enhances HDR-mediated gene repair with customizable endonucleases

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Diane Yang ◽  
Marissa A Scavuzzo ◽  
Jolanta Chmielowiec ◽  
Robert Sharp ◽  
Aleksandar Bajic ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Human Pluripotent Stem Cells ◽  
Cell Cycle Phase ◽  
Cycle Phase ◽  
Gene Repair ◽  
M Cell
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