Electrophysiological recordings of cardiomyocytes isolated from engineered human cardiac tissues derived from pluripotent stem cells

2018 ◽  
Author(s):  
Kumi Morikawa ◽  
LouJin Song ◽  
Kacey Ronaldson-Bouchard ◽  
Gordana Vunjak-Novakovic ◽  
Masayuki Yazawa
Keyword(s):  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Cardiac Tissues ◽  
Electrophysiological Recordings

scholarly journals Optimized Protocol to Generate Spinal Motor Neuron Cells from Induced Pluripotent Stem Cells from Charcot Marie Tooth Patients

2020 ◽  
Vol 10 (7) ◽  
pp. 407
Author(s):  
Pierre-Antoine Faye ◽  
Nicolas Vedrenne ◽  
Federica Miressi ◽  
Marion Rassat ◽  
Sergii Romanenko ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Motor Neurons ◽  
Pluripotent Stem Cells ◽  
Charcot Marie Tooth ◽  
Charcot Marie Tooth Disease ◽  
Spinal Motor ◽  
Electrophysiological Recordings ◽  
Induced Pluripotent

Modelling rare neurogenetic diseases to develop new therapeutic strategies is highly challenging. The use of human-induced pluripotent stem cells (hiPSCs) is a powerful approach to obtain specialized cells from patients. For hereditary peripheral neuropathies, such as Charcot–Marie–Tooth disease (CMT) Type II, spinal motor neurons (MNs) are impaired but are very difficult to study. Although several protocols are available to differentiate hiPSCs into neurons, their efficiency is still poor for CMT patients. Thus, our goal was to develop a robust, easy, and reproducible protocol to obtain MNs from CMT patient hiPSCs. The presented protocol generates MNs within 20 days, with a success rate of 80%, using specifically chosen molecules, such as Sonic Hedgehog or retinoic acid. The timing and concentrations of the factors used to induce differentiation are crucial and are given hereby. We then assessed the MNs by optic microscopy, immunocytochemistry (Islet1/2, HB9, Tuj1, and PGP9.5), and electrophysiological recordings. This method of generating MNs from CMT patients in vitro shows promise for the further development of assays to understand the pathological mechanisms of CMT and for drug screening.

scholarly journals Tubular Cardiac Tissues Derived from Human Induced Pluripotent Stem Cells Generate Pulse Pressure In Vivo

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Hiroyoshi Seta ◽  
Katsuhisa Matsuura ◽  
Hidekazu Sekine ◽  
Kenji Yamazaki ◽  
Tatsuya Shimizu
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pulse Pressure ◽  
Pluripotent Stem Cells ◽  
Cardiac Tissues ◽  
Induced Pluripotent

scholarly journals Bioengineering Clinically Relevant Cardiomyocytes and Cardiac Tissues from Pluripotent Stem Cells

2021 ◽  
Vol 22 (6) ◽  
pp. 3005
Author(s):  
Emma Claire James ◽  
Eva Tomaskovic-Crook ◽  
Jeremy Micah Crook
Keyword(s):  
Stem Cells ◽  
Heart Disease ◽  
Electric Fields ◽  
Pluripotent Stem Cells ◽  
Heart Development ◽  
Disease Patient ◽  
Toxicity Testing ◽  
Model Systems ◽  
Patient Specific ◽  
Cardiac Tissues

The regenerative capacity of cardiomyocytes is insufficient to functionally recover damaged tissue, and as such, ischaemic heart disease forms the largest proportion of cardiovascular associated deaths. Human-induced pluripotent stem cells (hiPSCs) have enormous potential for developing patient specific cardiomyocytes for modelling heart disease, patient-based cardiac toxicity testing and potentially replacement therapy. However, traditional protocols for hiPSC-derived cardiomyocytes yield mixed populations of atrial, ventricular and nodal-like cells with immature cardiac properties. New insights gleaned from embryonic heart development have progressed the precise production of subtype-specific hiPSC-derived cardiomyocytes; however, their physiological immaturity severely limits their utility as model systems and their use for drug screening and cell therapy. The long-entrenched challenges in this field are being addressed by innovative bioengingeering technologies that incorporate biophysical, biochemical and more recently biomimetic electrical cues, with the latter having the potential to be used to both direct hiPSC differentiation and augment maturation and the function of derived cardiomyocytes and cardiac tissues by mimicking endogenous electric fields.

scholarly journals Microglia-like Cells Promote Neuronal Functions in Cerebral Organoids

Cells ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 124
Author(s):  
Ilkka Fagerlund ◽  
Antonios Dougalis ◽  
Anastasia Shakirzyanova ◽  
Mireia Gómez-Budia ◽  
Anssi Pelkonen ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Synaptic Activity ◽  
Repetitive Firing ◽  
Electrophysiological Recordings ◽  
Induced Pluripotent ◽  
Neuronal Functions

Human cerebral organoids, derived from induced pluripotent stem cells, offer a unique in vitro research window to the development of the cerebral cortex. However, a key player in the developing brain, the microglia, do not natively emerge in cerebral organoids. Here we show that erythromyeloid progenitors (EMPs), differentiated from induced pluripotent stem cells, migrate to cerebral organoids, and mature into microglia-like cells and interact with synaptic material. Patch-clamp electrophysiological recordings show that the microglia-like population supported the emergence of more mature and diversified neuronal phenotypes displaying repetitive firing of action potentials, low-threshold spikes and synaptic activity, while multielectrode array recordings revealed spontaneous bursting activity and increased power of gamma-band oscillations upon pharmacological challenge with NMDA. To conclude, microglia-like cells within the organoids promote neuronal and network maturation and recapitulate some aspects of microglia-neuron co-development in vivo, indicating that cerebral organoids could be a useful biorealistic human in vitro platform for studying microglia-neuron interactions.

scholarly journals 3094 Production of Engineered Cardiac Tissue for Disease Modeling

2019 ◽  
Vol 3 (s1) ◽  
pp. 18-19
Author(s):  
Morgan Ellis ◽  
Elizabeth Lipke
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Pluripotent Stem Cells ◽  
Cardiac Tissue ◽  
Disease Modeling ◽  
Cardiac Regeneration ◽  
Cardiac Markers ◽  
Cardiac Tissues ◽  
Over Time

OBJECTIVES/SPECIFIC AIMS: Cardiovascular diseases (CVD) is the leading cause of death worldwide in both men and women due to lack of cardiac regeneration after disease or damaged is caused. There are many challenges to studying CVD since native cardiomyocytes cannot be cultured in vitro. With the advancements in biomaterial and pluripotent stem cells research, scientists are now able to produce engineered cardiac tissue models in vitro that mimic the native myocardium. This study shows our methods for producing engineered cardiac tissue with potential applications in cardiac regeneration, disease modeling, and scalable production. METHODS/STUDY POPULATION: In this study, human induced pluripotent stem cells (hiPSCs) were combined with two different photocrosslinkable hybrid biomaterials, poly (ethylene)- glycol fibrinogen (PF) and gelatin methacrylate (GelMa), in various tissue geometries to form 3D human engineered cardiac tissues (3D-hECTs). To study tissue growth and contraction, image and video analysis was performed at specific timepoints. To analyze differentiation efficiency and cell population, flow cytometry was performed using cardiac markers. To evaluate gene expression, qPCR was performed using pluripotency and cardiac specific primers. RESULTS/ANTICIPATED RESULTS: Direct cardiac differentiation of encapsulated hiPSCs resulted in synchronously contracting 3D-hECTs in both biomaterials and all tissue geometries. Spontaneous contractions started on Day 7 and increased in velocity, frequency, and synchronicity over time. 3D-hECTs had high cell viability with > 70% of cells positive for cardiac markers. Engineered tissues showed appropriate temporal changes in gene expression over time with pluripotency gene expression decreasing and cardiac gene expression increasing. DISCUSSION/SIGNIFICANCE OF IMPACT: This study shows the potential for direct differentiation of encapsulated hiPSCs to produce physiologically relevant engineered cardiac tissues. Resulting 3D-hECTS showed features of mature myocardium with appropriate cardiomyocyte populations, mechanical motion, and gene expression. Using this platform, we are able to produce engineered cardiac tissue in a variety of biomaterials and tissue geometries to study new therapeutics, mechanism of disease, and scalable tissue culture.

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