MitoCellPhe reveals mitochondrial morphologies in single fibroblasts and clustered stem cells

2021 ◽  
Author(s):  
Ajibola B. Bakare ◽  
Fibi Meshrkey ◽  
Benjamin Lowe ◽  
Carson Molder ◽  
Raj R Rao ◽  
...  
Keyword(s):  
Stem Cell ◽  
Mitochondrial Dynamics ◽  
Single Cells ◽  
Biological Significance ◽  
Induced Pluripotent Stem Cell ◽  
Fibroblast Cell ◽  
Cell Types ◽  
Health And Disease ◽  
Diseased State ◽  
Induced Pluripotent

Mitochondria are dynamic organelles that differ significantly in their morphologies across cell types, reflecting specific cellular needs and stages in development. Despite the wide biological significance in disease and health, delineating mitochondrial morphologies in complex systems remains challenging. Here, we present the Mitochondrial Cellular Phenotype (MitoCellPhe) tool developed for quantifying mitochondrial morphologies and demonstrate its utility in delineating differences in mitochondrial morphologies in a human fibroblast and human induced pluripotent stem cell (hiPSC) line. MitoCellPhe generates 24 parameters, allowing for a comprehensive analysis of mitochondrial structures and importantly allows for quantification to be performed on mitochondria in images containing single cells or clusters of cells. With this tool, we were able to validate previous findings that show networks of mitochondria in healthy fibroblast cell lines and a more fragmented morphology in hiPSCs. Using images generated from control and diseased fibroblasts and hiPSCs, we also demonstrate the efficacy of the toolset in delineating differences in morphologies between healthy and the diseased state in both stem cell (hiPSC) and differentiated fibroblast cells. Our results demonstrate that MitoCellPhe enables high-throughput, sensitive, detailed and quantitative mitochondrial morphological assessment and thus enables better biological insights into mitochondrial dynamics in health and disease

scholarly journals Induced pluripotent stem cell technology for modelling and therapy of cerebellar ataxia

Open Biology ◽  
2015 ◽  
Vol 5 (7) ◽  
pp. 150056 ◽  
Author(s):  
Lauren M. Watson ◽  
Maggie M. K. Wong ◽  
Esther B. E. Becker
Keyword(s):  
Stem Cell ◽  
Cerebellar Ataxia ◽  
Pluripotent Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
Cell Types ◽  
Disease Modelling ◽  
Stem Cell Technology ◽  
Induced Pluripotent ◽  
First Descriptions ◽  
Made In

Induced pluripotent stem cell (iPSC) technology has emerged as an important tool in understanding, and potentially reversing, disease pathology. This is particularly true in the case of neurodegenerative diseases, in which the affected cell types are not readily accessible for study. Since the first descriptions of iPSC-based disease modelling, considerable advances have been made in understanding the aetiology and progression of a diverse array of neurodegenerative conditions, including Parkinson's disease and Alzheimer's disease. To date, however, relatively few studies have succeeded in using iPSCs to model the neurodegeneration observed in cerebellar ataxia. Given the distinct neurodevelopmental phenotypes associated with certain types of ataxia, iPSC-based models are likely to provide significant insights, not only into disease progression, but also to the development of early-intervention therapies. In this review, we describe the existing iPSC-based disease models of this heterogeneous group of conditions and explore the challenges associated with generating cerebellar neurons from iPSCs, which have thus far hindered the expansion of this research.

scholarly journals Mitochondrial dysfunction in iPSC-derived neurons of subjects with chronic mountain sickness

2018 ◽  
Vol 125 (3) ◽  
pp. 832-840 ◽  
Author(s):  
Helen Zhao ◽  
Guy Perkins ◽  
Hang Yao ◽  
David Callacondo ◽  
Otto Appenzeller ◽  
...  
Keyword(s):  
Cell Death ◽  
Stem Cell ◽  
Pluripotent Stem Cell ◽  
Mitochondrial Dynamics ◽  
Induced Pluripotent Stem Cell ◽  
Mitochondrial Morphology ◽  
Chronic Mountain Sickness ◽  
Atp Levels ◽  
Mountain Sickness ◽  
Induced Pluripotent

Patients with chronic mountain sickness (CMS) suffer from hypoxemia, erythrocytosis, and numerous neurologic deficits. Here we used induced pluripotent stem cell (iPSC)-derived neurons from both CMS and non-CMS subjects to study CMS neuropathology. Using transmission electron microscopy, we report that CMS neurons have a decreased mitochondrial volume density, length, and less cristae membrane surface area. Real-time PCR confirmed a decreased mitochondrial fusion gene optic atrophy 1 (OPA1) expression. Immunoblot analysis showed an accumulation of the short isoform of OPA1 (S-OPA1) in CMS neurons, which have reduced ATP levels under normoxia and increased lactate dehydrogenase (LDH) release and caspase 3 activation after hypoxia. Improving the balance between the long isoform of OPA1 and S-OPA1 in CMS neurons increased the ATP levels and attenuated LDH release under hypoxia. Our data provide initial evidence for altered mitochondrial morphology and function in CMS neurons, and reveal increased cell death under hypoxia due in part to altered mitochondrial dynamics. NEW & NOTEWORTHY Induced pluripotent stem cell-derived neurons from chronic mountain sickness (CMS) subjects have altered mitochondrial morphology and dynamics, and increased sensitivity to hypoxic stress. Modification of OPA1 can attenuate cell death after hypoxic treatment, providing evidence that altered mitochondrial dynamics play an important role in increased vulnerability under stress in CMS neurons.

scholarly journals Reduction of HDAC2 expression in human induced pluripotent stem cell derived neurons improves neuronal maturation, mitochondrial dynamics and cellular neurodegenerative disease phenotypes

2021 ◽  
Author(s):  
Harald Frankowski ◽  
Fred Yeboah ◽  
Bonnie J. Berry ◽  
Chizuru Kinoshita ◽  
Michelle Lee ◽  
...  
Keyword(s):  
Stem Cell ◽  
Neurodegenerative Disease ◽  
Pluripotent Stem Cell ◽  
Mitochondrial Dynamics ◽  
Induced Pluripotent Stem Cell ◽  
Adult Brain ◽  
Aberrant Expression ◽  
Histone Deacetylase 2 ◽  
Knock Down ◽  
Induced Pluripotent

AbstractHistone deacetylase 2 (HDAC2) is a major HDAC protein in the adult brain and has been shown to regulate many neuronal genes. Aberrant expression of HDAC2 and subsequent dysregulation of neuronal gene expression is implicated in neurodegeneration and brain aging. Human induced pluripotent stem cell-derived neurons (hiPSC-Ns) are widely used models for studying neurodegenerative disease mechanisms, but the role of HDAC2 in hiPSC-N differentiation and maturation has not been explored. In this study, we show that levels of HDAC2 progressively decrease as hiPSCs are differentiated towards neurons. This suppression of HDAC2 inversely corresponds to an increase in neuron-specific isoforms of Endophilin-B1, a multifunctional protein involved in mitochondrial dynamics. Expression of neuron-specific isoforms of Endophilin-B1 is accompanied by concomitant expression of a neuron-specific alternative splicing factor, SRRM4. Manipulation of HDAC2 and Endophilin-B1 using lentiviral approaches shows that knock-down of HDAC2 or overexpression of a neuron-specific Endophilin-B1 isoform promotes mitochondrial elongation and protects against cytotoxic stress in hiPSC-Ns, while HDAC2 knock-down specifically influences genes regulating mitochondrial dynamics and synaptogenesis. Furthermore, HDAC2 knock-down promotes enhanced mitochondrial respiration and reduces levels of neurotoxic amyloid beta peptides. Collectively, our study demonstrates a role for HDAC2 in hiPSC-neuronal differentiation, highlights neuron-specific isoforms of Endophilin-B1 as a marker of differentiating hiPSC-Ns, and demonstrates that HDAC2 regulates key neuronal and mitochondrial pathways in hiPSC-Ns.

Stem Cells: In Sickness and in Health

2020 ◽  
Vol 15 ◽  
Author(s):  
Hisham F. Bahmada ◽  
Mohamad K. Elajami ◽  
Reem Daouk ◽  
Hiba Jalloul ◽  
Batoul Darwish ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
Cell Types ◽  
Therapy Resistance ◽  
The Body ◽  
Differentiated Cells ◽  
Undifferentiated Cells ◽  
Potential Applications ◽  
Induced Pluripotent

: Stem cells are undifferentiated cells with the ability to proliferate and convert to different types of differentiated cells that make up the various tissues and organs in the body. They exist both in embryos as pluripotent stem cells that can differentiate into the three germ layers and as multipotent or unipotent stem cells in adult tissues to aid in repair and homeostasis. Perturbations in these cells’ normal functions can give rise to a wide variety of diseases. In this review, we discuss the origin of different stem cell types, their properties and characteristics, their role in tissue homeostasis, current research, and their potential applications in various life-threatening diseases. We focus on neural stem cells, their role in neurogenesis and how they can be exploited to treat diseases of the brain including neurodegenerative diseases and cancer. Next, we explore current research in induced pluripotent stem cell (iPSC) techniques and their clinical applications in regenerative and personalized medicine. Lastly, we tackle a special type of stem cells called cancer stem cells (CSCs) and how they can be responsible for therapy resistance and tumor recurrence and explore ways to target them.

scholarly journals Multimodal on-axis platform for all-optical electrophysiology with near-infrared probes in human stem-cell-derived cardiomyocytes

2018 ◽  
Author(s):  
Aleksandra Klimas ◽  
Gloria Ortiz ◽  
Steven Boggess ◽  
Evan W. Miller ◽  
Emilia Entcheva
Keyword(s):  
Stem Cell ◽  
High Throughput ◽  
High Performance ◽  
Near Infrared ◽  
Induced Pluripotent Stem Cell ◽  
Cell Types ◽  
Simultaneous Recording ◽  
Optogenetic Stimulation ◽  
All Optical ◽  
Induced Pluripotent

AbstractCombined optogenetic stimulation and optical imaging permits scalable, high-throughput probing of cellular electrophysiology and optimization of stem-cell derived excitable cells, such as neurons and muscle cells. We report a new “on-axis” configuration of OptoDyCE, our all-optical platform for studying human induced pluripotent stem-cell-derived cardiomyocytes (hiPSC-CMs) and other cell types, optically driven by Channelrhodopsin2 (ChR2). This solid-state system integrates optogenetic stimulation with temporally-multiplexed simultaneous recording of membrane voltage (Vm) and intracellular calcium ([Ca2+]i) dynamics using a single photodetector. We demonstrate the capacity for combining multiple spectrally-compatible actuators and sensors, including newer high-performance near-infrared (NIR) voltage probes BeRST1 and Di-4-ANBDQBS, to record complex spatiotemporal responses of hiPSC-CMs to drugs in a high-throughput manner.

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