scholarly journals A chemical tool for improved culture of human pluripotent stem cells

2020 ◽  
Author(s):  
Laurence Silpa ◽  
Maximilian Schuessler ◽  
Gu Liu ◽  
Marcus Olivecrona ◽  
Lucia Groizard-Payeras ◽  
...  
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Large Scale ◽  
3D Culture ◽  
Cost Effective ◽  
Human Pluripotent Stem Cells ◽  
Directed Differentiation ◽  
Kinase C ◽  
2D And 3D ◽  
Cell Cell

AbstractThe large-scale and cost-effective production of quality-controlled human pluripotent stem cells (hPSC) for use in cell therapy and drug discovery requires chemically-defined xenobiotic-free culture systems that enable easy and homogeneous expansion of pluripotent cells. Through phenotypic screening, we have identified a small molecule, OXS8360 (an optimized derivative of (-)-Indolactam V ((-)-ILV)), that stably disrupts hPSC cell-cell contacts. Proliferation of hPSC in OXS8360 is normal, as are pluripotency signatures, directed differentiation to hallmark lineages and karyotype over extended passaging. In 3D culture, OXS8360-treated hPSC form smaller, more uniform aggregates, that are easier to dissociate, greatly facilitating expansion. The mode of action of OXS8360 involves disruption of the localisation of the cell-cell adhesion molecule E-cadherin, via activation of unconventional Protein Kinase C isoforms. OXS8360 media supplementation is therefore able to yield more uniform, disaggregated 2D and 3D hPSC cultures, providing the hPSC field with an affordable tool to improve hPSC quality and scalability.

scholarly journals Characterization of Phenotypic and Transcriptional Differences in Human Pluripotent Stem Cells under 2D and 3D Culture Conditions

2016 ◽  
Vol 5 (22) ◽  
pp. 2951-2958 ◽  
Author(s):  
Ken-ichiro Kamei ◽  
Yoshie Koyama ◽  
Yumie Tokunaga ◽  
Yasumasa Mashimo ◽  
Momoko Yoshioka ◽  
...  
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
3D Culture ◽  
Culture Conditions ◽  
Human Pluripotent Stem Cells ◽  
2D And 3D

scholarly journals Assessment of the Hematopoietic Differentiation Potential of Human Pluripotent Stem Cells in 2D and 3D Culture Systems

Cells ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 2858
Author(s):  
German Atzin Mora-Roldan ◽  
Dalia Ramirez-Ramirez ◽  
Rosana Pelayo ◽  
Karlen Gazarian
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Time Course ◽  
Hematopoietic Cells ◽  
3D Culture ◽  
Human Pluripotent Stem Cells ◽  
Hematopoietic Differentiation ◽  
Differentiation Potential ◽  
2D And 3D

Background. In vitro methods for hematopoietic differentiation of human pluripotent stem cells (hPSC) are a matter of priority for the in-depth research into the mechanisms of early embryogenesis. So-far, published results regarding the generation of hematopoietic cells come from studies using either 2D or 3D culture formats, hence, it is difficult to discern their particular contribution to the development of the concept of a unique in vitro model in close resemblance to in vivo hematopoiesis. Aim of the study. To assess using the same culture conditions and the same time course, the potential of each of these two formats to support differentiation of human pluripotent stem cells to primitive hematopoiesis without exogenous activation of Wnt signaling. Methods. We used in parallel 2D and 3D formats, the same culture environment and assay methods (flow cytometry, IF, qPCR) to investigate stages of commitment and specification of mesodermal, and hemogenic endothelial cells to CD34 hematopoietic cells and evaluated their clonogenic capacity in a CFU system. Results. We show an adequate formation of mesoderm, an efficient commitment to hemogenic endothelium, a higher number of CD34 hematopoietic cells, and colony-forming capacity potential only in the 3D format-supported differentiation. Conclusions. This study shows that the 3D but not the 2D format ensures the induction and realization by endogenous mechanisms of human pluripotent stem cells’ intrinsic differentiation program to primitive hematopoietic cells. We propose that the 3D format provides an adequate level of upregulation of the endogenous Wnt/β-catenin signaling.

scholarly journals Directed differentiation and long-term maintenance of epicardial cells derived from human pluripotent stem cells under fully defined conditions

2017 ◽  
Vol 12 (9) ◽  
pp. 1890-1900 ◽  
Author(s):  
Xiaoping Bao ◽  
Xiaojun Lian ◽  
Tongcheng Qian ◽  
Vijesh J Bhute ◽  
Tianxiao Han ◽  
...  
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Human Pluripotent Stem Cells ◽  
Directed Differentiation ◽  
Epicardial Cells ◽  
Term Maintenance

scholarly journals Modulating cell state to enhance suspension expansion of human pluripotent stem cells

2018 ◽  
Vol 115 (25) ◽  
pp. 6369-6374 ◽  
Author(s):  
Yonatan Y. Lipsitz ◽  
Curtis Woodford ◽  
Ting Yin ◽  
Jacob H. Hanna ◽  
Peter W. Zandstra
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Large Scale ◽  
Cell Types ◽  
Human Pluripotent Stem Cells ◽  
The Body ◽  
Altered Expression ◽  
Pancreatic Progenitors ◽  
Erk Inhibition

The development of cell-based therapies to replace missing or damaged tissues within the body or generate cells with a unique biological activity requires a reliable and accessible source of cells. Human pluripotent stem cells (hPSC) have emerged as a strong candidate cell source capable of extended propagation in vitro and differentiation to clinically relevant cell types. However, the application of hPSC in cell-based therapies requires overcoming yield limitations in large-scale hPSC manufacturing. We explored methods to convert hPSC to alternative states of pluripotency with advantageous bioprocessing properties, identifying a suspension-based small-molecule and cytokine combination that supports increased single-cell survival efficiency, faster growth rates, higher densities, and greater expansion than control hPSC cultures. ERK inhibition was found to be essential for conversion to this altered state, but once converted, ERK inhibition led to a loss of pluripotent phenotype in suspension. The resulting suspension medium formulation enabled hPSC suspension yields 5.7 ± 0.2-fold greater than conventional hPSC in 6 d, for at least five passages. Treated cells remained pluripotent, karyotypically normal, and capable of differentiating into all germ layers. Treated cells could also be integrated into directed differentiated strategies as demonstrated by the generation of pancreatic progenitors (NKX6.1+/PDX1+ cells). Enhanced suspension-yield hPSC displayed higher oxidative metabolism and altered expression of adhesion-related genes. The enhanced bioprocess properties of this alternative pluripotent state provide a strategy to overcome cell manufacturing limitations of hPSC.

scholarly journals Functional cortical neurons and astrocytes from human pluripotent stem cells in 3D culture

2015 ◽  
Vol 12 (7) ◽  
pp. 671-678 ◽  
Author(s):  
Anca M Paşca ◽  
Steven A Sloan ◽  
Laura E Clarke ◽  
Yuan Tian ◽  
Christopher D Makinson ◽  
...  
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Cortical Neurons ◽  
3D Culture ◽  
Human Pluripotent Stem Cells
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