scholarly journals Circulating re-entrant waves promote maturation of hiPSC-derived cardiomyocytes in self-organized tissue ring

2020 ◽  
Vol 3 (1) ◽  
Author(s):  
Junjun Li ◽  
Lu Zhang ◽  
Leqian Yu ◽  
Itsunari Minami ◽  
Shigeru Miyagawa ◽  
...  
Keyword(s):  
Pluripotent Stem Cells ◽  
Consumption Rate ◽  
Three Dimensional ◽  
Specific Gene ◽  
Directed Differentiation ◽  
Self Organized ◽  
Induced Pluripotent ◽  
Electrical Pacing ◽  
Long Term Behavior

AbstractDirected differentiation methods allow acquisition of high-purity cardiomyocytes differentiated from human induced pluripotent stem cells (hiPSCs); however, their immaturity characteristic limits their application for drug screening and regenerative therapy. The rapid electrical pacing of cardiomyocytes has been used for efficiently promoting the maturation of cardiomyocytes, here we describe a simple device in modified culture plate on which hiPSC-derived cardiomyocytes can form three-dimensional self-organized tissue rings (SOTRs). Using calcium imaging, we show that within the ring, reentrant waves (ReWs) of action potential spontaneously originated and ran robustly at a frequency up to 4 Hz. After 2 weeks, SOTRs with ReWs show higher maturation including structural organization, increased cardiac-specific gene expression, enhanced Ca2+-handling properties, an increased oxygen-consumption rate, and enhanced contractile force. We subsequently use a mathematical model to interpret the origination, propagation, and long-term behavior of the ReWs within the SOTRs.

scholarly journals Rapid pacing by circulating traveling waves improves maturation of hiPSC-derived cardiomyocytes in self-organized tissue ring

2019 ◽  
Author(s):  
Junjun Li ◽  
Lu Zhang ◽  
Leqian Yu ◽  
Itsunari Minami ◽  
Marcel Hörning ◽  
...  
Keyword(s):  
Traveling Waves ◽  
Three Dimensional ◽  
Specific Gene ◽  
Excitable Cells ◽  
Self Organized ◽  
Rapid Pacing ◽  
Induced Pluripotent ◽  
Electrical Pacing ◽  
Long Term Behavior

AbstractDirected differentiation methods allow acquisition of high-purity cardiomyocytes (CMs) differentiated from human induced pluripotent stem cells (hiPSCs); however, their immaturity characteristic limits their application for drug screening and regenerative therapy. The rapid electrical pacing of cardiomyocytes have been used for efficiently promoting the maturation of cardiomyocytes, here we describe a simple device in modified culture plate on which hiPSC-derived CMs (hiPSC-CMs) can form three-dimensional self-organized tissue rings (SOTRs). Using calcium imaging, we show that within the ring, traveling waves (TWs) of action potential spontaneously originated and ran robustly at a frequency up to 4 Hz. After 2 weeks, SOTRs with TW training showed matured features including structural organization, increased cardiac-specific gene expression, enhanced Ca2+-handling properties, an increased oxygen-consumption rate, and enhanced contractile force. We subsequently used a mathematical model to interpret the origination, propagation, and long-term behavior of the TWs within the SOTRs. This new idea for spontaneous hiPSC-CM maturation also has potential for pacing the electrical excitable cells such as neuron and retina cells for various applications.

scholarly journals Long-Term Stability and Differentiation Potential of Cryopreserved cGMP-Compliant Human Induced Pluripotent Stem Cells

2019 ◽  
Vol 21 (1) ◽  
pp. 108 ◽  
Author(s):  
Mehdi Shafa ◽  
Tylor Walsh ◽  
Krishna Morgan Panchalingam ◽  
Thomas Richardson ◽  
Laura Menendez ◽  
...  
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Genomic Stability ◽  
Directed Differentiation ◽  
Differentiation Potential ◽  
Long Term Stability ◽  
Induced Pluripotent ◽  
Cell Banks ◽  
2D And 3D

The clinical effectiveness of human induced pluripotent stem cells (iPSCs) is highly dependent on a few key quality characteristics including the generation of high quality cell bank, long-term genomic stability, post-thaw viability, plating efficiency, retention of pluripotency, directed differentiation, purity, potency, and sterility. We have already reported the establishment of iPSC master cell banks (MCBs) and working cell banks (WCBs) under current good manufacturing procedure (cGMP)-compliant conditions. In this study, we assessed the cellular and genomic stability of the iPSC lines generated and cryopreserved five years ago under cGMP-compliant conditions. iPSC lines were thawed, characterized, and directly differentiated into cells from three germ layers including cardiomyocytes (CMs), neural stem cells (NSCs), and definitive endoderm (DE). The cells were also expanded in 2D and 3D spinner flasks to evaluate their long-term expansion potential in matrix-dependent and feeder-free culture environment. All three lines successfully thawed and attached to the L7TM matrix, and formed typical iPSC colonies that expressed pluripotency markers over 15 passages. iPSCs maintained their differentiation potential as demonstrated with spontaneous and directed differentiation to the three germ layers and corresponding expression of specific markers, respectfully. Furthermore, post-thaw cells showed normal karyotype, negative mycoplasma, and sterility testing. These cells maintained both their 2D and 3D proliferation potential after five years of cryopreservation without acquiring karyotype abnormality, loss of pluripotency, and telomerase activity. These results illustrate the long-term stability of cGMP iPSC lines, which is an important step in establishing a reliable, long-term source of starting materials for clinical and commercial manufacturing of iPSC-derived cell therapy products.

scholarly journals Cell Reprogramming, IPS Limitations, and Overcoming Strategies in Dental Bioengineering

2012 ◽  
Vol 2012 ◽  
pp. 1-8 ◽  
Author(s):  
Gaskon Ibarretxe ◽  
Antonia Alvarez ◽  
Maria-Luz Cañavate ◽  
Enrique Hilario ◽  
Maitane Aurrekoetxea ◽  
...  
Keyword(s):  
Pluripotent Stem Cells ◽  
Ips Cells ◽  
Cell Reprogramming ◽  
Animal Cells ◽  
Ips Cell ◽  
Current State ◽  
Organ Systems ◽  
Induced Pluripotent ◽  
Long Term Behavior

The procurement of induced pluripotent stem cells, or IPS cells, from adult differentiated animal cells has the potential to revolutionize future medicine, where reprogrammed IPS cells may be used to repair disease-affected tissues on demand. The potential of IPS cell technology is tremendous, but it will be essential to improve the methodologies for IPS cell generation and to precisely evaluate each clone and subclone of IPS cells for their safety and efficacy. Additionally, the current state of knowledge on IPS cells advises that research on their regenerative properties is carried out in appropriate tissue and organ systems that permit a safe assessment of the long-term behavior of these reprogrammed cells. In the present paper, we discuss the mechanisms of cell reprogramming, current technical limitations of IPS cells for their use in human tissue engineering, and possibilities to overcome them in the particular case of dental regeneration.

Enhanced microvasculature formation and patterning in iPSC-derived kidney organoids cultured in physiological hypoxia

2021 ◽  
Author(s):  
Anika Schumacher ◽  
Nadia Roumans ◽  
Timo Rademakers ◽  
Virginie Joris ◽  
Maria Jose Eischen-Loges ◽  
...  
Keyword(s):  
Pluripotent Stem Cells ◽  
Three Dimensional ◽  
Culture Conditions ◽  
End Stage Kidney Disease ◽  
Whole Mount ◽  
Factor Expression ◽  
Induced Pluripotent ◽  
End Stage ◽  
Kidney Organoids

Functional kidney organoids have the potential to be used in implantable kidney grafts for patients with end-stage kidney disease, because they have been shown to self-organize from induced pluripotent stem cells into most important renal structures. To date, however, long-term kidney organoid culture has not succeeded, as nephrons lose their phenotype after approximately 25 days. Furthermore, the renal structures remain immature with diminishing endothelial networks with low connectivity and limited organoid invasion. We hypothesized that introducing long-term culture at physiological hypoxia, rather than the normally applied non-physiological, hyperoxic 21% O2, could initiate angiogenesis, lead to enhanced growth factor expression and improve the endothelial patterning. We therefore cultured the kidney organoids at 7% O2 instead of 21% O2 for up to 25 days and evaluated nephrogenesis, VEGF-A expression and vascularization. Whole mount imaging revealed a homogenous morphology of the endothelial network with enhanced sprouting and interconnectivity when the kidney organoids were cultured in hypoxia. Three-dimensional quantification confirmed that the hypoxic culture led to an increased average vessel length, likely due to the observed upregulation of proangiogenic VEGF-A189 mRNA and downregulation of the antiangiogenic protein VEGF-A165b measured in hypoxia. This research indicates the importance of optimization of oxygen availability in organoid systems and the potential of hypoxic culture conditions in improving the vascularization of organoids.

Directed differentiation of mouse-induced pluripotent stem cells generates dopaminergic nerve

2010 ◽  
Vol 34 (8) ◽  
pp. S36-S36
Author(s):  
Ping Duan ◽  
Xuelin Ren ◽  
Wenhai Yan ◽  
Xuefei Han ◽  
Xu Yan ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Directed Differentiation ◽  
Induced Pluripotent

scholarly journals Chemically defined and xeno-free culture condition for human extended pluripotent stem cells

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Bei Liu ◽  
Shi Chen ◽  
Yaxing Xu ◽  
Yulin Lyu ◽  
Jinlin Wang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Culture Condition ◽  
Human Fibroblast ◽  
Culture System ◽  
Disease Modeling ◽  
Directed Differentiation

AbstractExtended pluripotent stem (EPS) cells have shown great applicative potentials in generating synthetic embryos, directed differentiation and disease modeling. However, the lack of a xeno-free culture condition has significantly limited their applications. Here, we report a chemically defined and xeno-free culture system for culturing and deriving human EPS cells in vitro. Xeno-free human EPS cells can be long-term and genetically stably maintained in vitro, as well as preserve their embryonic and extraembryonic developmental potentials. Furthermore, the xeno-free culturing system also permits efficient derivation of human EPS cells from human fibroblast through reprogramming. Our study could have broad utility in future applications of human EPS cells in biomedicine.

scholarly journals Novel hybrid three-dimensional artificial liver using human induced pluripotent stem cells and a rat decellularized liver scaffold

2019 ◽  
Vol 10 ◽  
pp. 127-133 ◽  
Author(s):  
Takahito Minami ◽  
Takamichi Ishii ◽  
Kentaro Yasuchika ◽  
Ken Fukumitsu ◽  
Satoshi Ogiso ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Three Dimensional ◽  
Artificial Liver ◽  
Induced Pluripotent ◽  
Decellularized Liver

scholarly journals Clonal genetic and hematopoietic heterogeneity among human-induced pluripotent stem cell lines

Blood ◽  
2013 ◽  
Vol 122 (12) ◽  
pp. 2047-2051 ◽  
Author(s):  
Jason A. Mills ◽  
Kai Wang ◽  
Prasuna Paluru ◽  
Lei Ying ◽  
Lin Lu ◽  
...  
Keyword(s):  
Pluripotent Stem Cells ◽  
Induced Pluripotent Stem Cell ◽  
Specific Gene ◽  
Rare Cnvs ◽  
Developmental Potential ◽  
Hematopoietic Development ◽  
Key Points ◽  
Stem Cell Lines ◽  
Induced Pluripotent ◽  
Selection Of

Key Points Normal induced pluripotent stem cells exhibit donor-specific gene expression signatures and the capacity for hematopoietic development. CNVs acquired during reprogramming or selection of rare CNVs present in the starting cell population may alter iPSC developmental potential.

scholarly journals Development of Multilayer Mesenchymal Stem Cell Cell Sheets

2021 ◽  
Vol 1 (1) ◽  
pp. 4-24
Author(s):  
Jun Ochiai ◽  
Yutaka Niihara ◽  
Joan Oliva
Keyword(s):  
Stem Cells ◽  
Stromal Cells ◽  
Pluripotent Stem Cells ◽  
Cell Sheets ◽  
Gene Therapies ◽  
The Past ◽  
Long Term Effect ◽  
Induced Pluripotent ◽  
New Guidelines

Cell and gene therapies have been developing dramatically over the past decade. To face and adapt to the development of these new therapies, the Food and Drug Administration (FDA) wrote and updated new guidelines from 2016 and keep updating them. Mesenchymal stem cells (MSCs) are the most used cells for treatment, far ahead from the induced pluripotent stem cells (iPSCs), based on registered clinical trials at clinicaltrials.gov. They are widely used because of their differentiation capacity and their anti-inflammatory properties, but some controversies still require clear answers. Additional studies are needed to determine the dosage, the number, and the route of injections (location and transplantation method), and if allogenic MSCs are safe compared to autologous MSC injection, including their long-term effect. In this review, we summarize the research our company is conducting with the adipose stromal cells in engineering cell sheets and their potential application.

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