scholarly journals Development of electrical activity in cardiac myocyte aggregates derived from mouse embryonic stem cells

2003 ◽  
Vol 284 (6) ◽  
pp. H2114-H2123 ◽  
Author(s):  
K. Banach ◽  
M. D. Halbach ◽  
P. Hu ◽  
J. Hescheler ◽  
U. Egert
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Action Potential ◽  
Electrical Activity ◽  
Cardiac Myocyte ◽  
Three Dimensional ◽  
Embryonic Stem ◽  
Electrophysiological Properties ◽  
Chronotropic Response

Embryonic stem cells differentiate into cardiac myocytes, repeating in vitro the structural and molecular changes associated with cardiac development. Currently, it is not clear whether the electrophysiological properties of the multicellular cardiac structure follow cardiac maturation as well. In long-term recordings of extracellular field potentials with microelectrode arrays consisting of 60 substrate-integrated electrodes, we examined the electrophysiological properties during the ongoing differentiation process. The beating frequency of the growing preparations increased from 1 to 5 Hz concomitant to a decrease of the action potential duration and action potential rise time. A developmental increase of the conduction velocity could be attributed to an increased expression of connexin43 gap junction channels. Whereas isoprenalin elicited a positive chronotropic response from the first day of spontaneous beating onward, a concentration-dependent negative chronotropic effect of carbachol only developed after ∼4 days. The in vitro development of the three-dimensional cardiac preparation thus closely follows the development described for the mouse embryonic heart, making it an ideal model to monitor the differentiation of electrical activity in embryonic cardiomyocytes.

scholarly journals Self-Assembling Scaffolds Supported Long-Term Growth of Human Primed Embryonic Stem Cells and Upregulated Core and Naïve Pluripotent Markers

Cells ◽  
2019 ◽  
Vol 8 (12) ◽  
pp. 1650 ◽  
Author(s):  
Christina McKee ◽  
Christina Brown ◽  
G. Rasul Chaudhry
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Three Dimensional ◽  
Embryonic Stem ◽  
Extended Period ◽  
Culture Conditions ◽  
Differentiation Potential ◽  
Self Assembling

The maintenance and expansion of human embryonic stem cells (ESCs) in two-dimensional (2-D) culture is technically challenging, requiring routine manipulation and passaging. We developed three-dimensional (3-D) scaffolds to mimic the in vivo microenvironment for stem cell proliferation. The scaffolds were made of two 8-arm polyethylene glycol (PEG) polymers functionalized with thiol (PEG-8-SH) and acrylate (PEG-8-Acr) end groups, which self-assembled via a Michael addition reaction. When primed ESCs (H9 cells) were mixed with PEG polymers, they were encapsulated and grew for an extended period, while maintaining their viability, self-renewal, and differentiation potential both in vitro and in vivo. Three-dimensional (3-D) self-assembling scaffold-grown cells displayed an upregulation of core pluripotency genes, OCT4, NANOG, and SOX2. In addition, the expression of primed markers decreased, while the expression of naïve markers substantially increased. Interestingly, the expression of mechanosensitive genes, YAP and TAZ, was also upregulated. YAP inhibition by Verteporfin abrogated the increased expression of YAP/TAZ as well as core and naïve pluripotent markers. Evidently, the 3-D culture conditions induced the upregulation of makers associated with a naïve state of pluripotency in the primed cells. Overall, our 3-D culture system supported the expansion of a homogenous population of ESCs and should be helpful in advancing their use for cell therapy and regenerative medicine.

Three‐dimensional electrospun gelatin scaffold coseeded with embryonic stem cells and sertoli cells: A promising substrate for in vitro coculture system

2019 ◽  
Vol 120 (8) ◽  
pp. 12508-12518 ◽  
Author(s):  
Mina Vardiani ◽  
Mazaher Gholipourmalekabadi ◽  
Marefat Ghaffari Novin ◽  
Morteza Koruji ◽  
Hatef Ghasemi Hamidabadi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Sertoli Cells ◽  
Three Dimensional ◽  
Embryonic Stem ◽  
Coculture System ◽  
Electrospun Gelatin

scholarly journals 3D synthetic microscaffolds promote homogenous expression of NANOG in mouse embryonic stem cells

2020 ◽  
Author(s):  
Natalia A. Bakhtina ◽  
Madlen Müller ◽  
Harry Wischnewski ◽  
Rajika Arora ◽  
Constance Ciaudo
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Three Dimensional ◽  
Embryonic Stem ◽  
Cell Communication ◽  
Early Embryo ◽  
Pluripotency Factors ◽  
Physiological Relevance ◽  
Synthetic Scaffold

The development of in vitro models, which accurately recapitulate early embryonic development, is one of the fundamental challenges in stem cell research. Most of the currently employed approaches involve the culture of embryonic stem cells (ESCs) on two-dimensional (2D) surfaces. However, the monolayer nature of these cultures does not permit cells to grow and proliferate in realistic three-dimensional (3D) microenvironments, as in an early embryo. In this paper, novel 3D synthetic scaffold arrays, fabricated by two-photon polymerization photolithography, are utilized to mimic tissue-specific architecture, enabling cell-to-matrix interaction and cell-to-cell communication in vitro. Mouse ESCs (mESCs) are able to grow and proliferate on these structures and maintain their pluripotent state. Furthermore, the 3D microscaffold arrays are integrated into a microscopy slide allowing the evaluation of the expression of key pluripotency factors at the single-cell level. Comparing 2D and 3D surfaces, mESCs grown in serum+LIF on 3D microscaffolds exhibit a stronger and more homogenous expression of NANOG and OCT4 pluripotency factors, than cells cultivated in 2i media, demonstrating that 3D microscaffolds capture naive pluripotency in vitro. Thus, the slide affords a novel and unique tool to model and study mammalian early development with greater physiological relevance than conventional 2D cultures.

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