Fluorescent probes as a tool for cell population tracking in spontaneously active neural networks derived from human pluripotent stem cells

Mäkinen, Meeri; Joki, Tiina; Ylä‐Outinen, Laura; Skottman, Heli; Narkilahti, Susanna; Äänismaa, Riikka

doi:10.1016/j.jneumeth.2013.02.019

Cited by 10 publications

(11 citation statements)

References 22 publications

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“…35,36,127 However, many different in vitro systems can be used with MEA, including human stem cells and 3D brain cultures. 37,128 MEAs have been used in several studies of pharmacological and toxicological responses and can be used to assess the functionality of a neuronal model. 35,38,125,128 Finally, glial maturation and function, such as glutamate uptake and transformation into glutamine by astrocytes and myelination of oligodendrocytes are often assessed by cell specific markers 53,129 or biochemical assays.…”

Section: Future Applications and Assays Of A Human Brain-on-a-chipmentioning

confidence: 99%

“…37,128 MEAs have been used in several studies of pharmacological and toxicological responses and can be used to assess the functionality of a neuronal model. 35,38,125,128 Finally, glial maturation and function, such as glutamate uptake and transformation into glutamine by astrocytes and myelination of oligodendrocytes are often assessed by cell specific markers 53,129 or biochemical assays. 130,131 Several of these assays are mainly developed and applied for traditional 2D cell models why many of them need to be optimized for the more complex 3D models and can represent a challenge for the more complex MPS.…”

Section: Future Applications and Assays Of A Human Brain-on-a-chipmentioning

confidence: 99%

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Biological and medical applications of a brain-on-a-chip

Pamies

Hartung

Högberg

2014

Exp Biol Med (Maywood)

103

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The desire to develop and evaluate drugs as potential countermeasures for biological and chemical threats requires test systems that can also substitute for the clinical trials normally crucial for drug development. Current animal models have limited predictivity for drug efficacy in humans as the large majority of drugs fails in clinical trials. We have limited understanding of the function of the central nervous system and the complexity of the brain, especially during development and neuronal plasticity. Simple in vitro systems do not represent physiology and function of the brain. Moreover, the difficulty of studying interactions between human genetics and environmental factors leads to lack of knowledge about the events that induce neurological diseases. Microphysiological systems (MPS) promise to generate more complex in vitro human models that better simulate the organ’s biology and function. MPS combine different cell types in a specific three-dimensional (3D) configuration to simulate organs with a concrete function. The final aim of these MPS is to combine different “organoids” to generate a human-on-a-chip, an approach that would allow studies of complex physiological organ interactions. The recent discovery of induced pluripotent stem cells (iPSCs) gives a range of possibilities allowing cellular studies of individuals with different genetic backgrounds (e.g., human disease models). Application of iPSCs from different donors in MPS gives the opportunity to better understand mechanisms of the disease and can be a novel tool in drug development, toxicology, and medicine. In order to generate a brain-on-a-chip, we have established a 3D model from human iPSCs based on our experience with a 3D rat primary aggregating brain model. After four weeks of differentiation, human 3D aggregates stain positive for different neuronal markers and show higher gene expression of various neuronal differentiation markers compared to 2D cultures. Here we present the applications and challenges of this emerging technology.

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Section: Future Applications and Assays Of A Human Brain-on-a-chipmentioning

confidence: 99%

Section: Future Applications and Assays Of A Human Brain-on-a-chipmentioning

confidence: 99%

Biological and medical applications of a brain-on-a-chip

Pamies

Hartung

Högberg

2014

Exp Biol Med (Maywood)

103

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“…However, only a few studies have described the direct differentiation of hES to highly active and stable neuronal networks. 15,31 Additionally, as a reason for insufficient gene silencing after retroviral reprogramming, the electrophysiological network maturation of advantageous hiPS cells (no ethical concerns, patient-specific disease modeling) was found to be even more complicated. 11 Furthermore, the generation of an intermediate progenitor population, which can be maintained and finally mature on demand in much shorter periods, is necessary to accelerate the extensive time course (more than two month) of functional hES/hiPS neuronal differentiation.…”

Section: Discussionmentioning

confidence: 99%

In vitro field potential monitoring on a multi-microelectrode array for the electrophysiological long-term screening of neural stem cell maturation

Seidel

Jahnke

Englich

et al. 2017

Analyst

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Due to the lack of appropriate cell models as well as automated electrophysiology monitoring technologies, the standardized identification of neurotoxic or protective effects in vitro remains a major problem in today's pharmaceutical ingredient development. Over the past few years, in vivo-like human pluripotent stem cell-derived neuronal networks have turned out to be a promising physiological cell source, if the establishment of robust and time-saving functional maturation strategies based on stable and expandable neural progenitor populations can be achieved. Here, we describe a multi-microelectrode array (MMEA)-based bioelectronics platform that was optimized for long-term electrophysiological activity monitoring of neuronal networks via field potential measurements. Differentiation of small molecule-based neuronal progenitors on MMEAs led to functional neurons within 15 days. More strikingly, these functional neuronal cultures could remain electrophysiologically stable on the MMEAs for more than four weeks. The observed electrophysiological properties correlated with the expression of typical neuron subtype markers and were further validated by specific neurotransmitter applications. With our established monitoring platform, we could show for the first time the long-term stability of the neural stem cell-like progenitor population to differentiate to electrophysiologically active dopaminergic neuronal networks for more than 80 passages. In conclusion, we provide a comprehensive long-term stable field potential monitoring platform based on stem cell-derived human neuronal networks that can be automated and up-scaled for standardized high-content screening applications e.g. in the field of neurotoxic and neuroprotective therapeutics identification.

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“…Several chemical probes have proven long term labeling of stem cells, including chloromethylfluorescein diacetate (CMFDA or CellTracker) and long chain carbocyanine dyes (like DiI, DiO, DiD, and CM-DiI; Sutton et al, 2008; Mäkinen et al, 2013; Boehm-Sturm et al, 2014). CMFDA and DiD labels were found to be stable for up to 4 weeks in human ES-cell derived neural cells in vitro (Mäkinen et al, 2013).…”

Section: Fluorescence Imagingmentioning

confidence: 99%

“…CMFDA and DiD labels were found to be stable for up to 4 weeks in human ES-cell derived neural cells in vitro (Mäkinen et al, 2013). Stem cell labeling prior to implantation is efficient but cell tracking is restricted to ex vivo fluorescence microscopy (Jablonska et al, 2010; Boehm-Sturm et al, 2014).…”

Section: Fluorescence Imagingmentioning

confidence: 99%

A review of novel optical imaging strategies of the stroke pathology and stem cell therapy in stroke

Aswendt

Adamczak

Tennstaedt

2014

Front. Cell. Neurosci.

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Transplanted stem cells can induce and enhance functional recovery in experimental stroke. Invasive analysis has been extensively used to provide detailed cellular and molecular characterization of the stroke pathology and engrafted stem cells. But post mortem analysis is not appropriate to reveal the time scale of the dynamic interplay between the cell graft, the ischemic lesion and the endogenous repair mechanisms. This review describes non-invasive imaging techniques which have been developed to provide complementary in vivo information. Recent advances were made in analyzing simultaneously different aspects of the cell graft (e.g., number of cells, viability state, and cell fate), the ischemic lesion (e.g., blood–brain-barrier consistency, hypoxic, and necrotic areas) and the neuronal and vascular network. We focus on optical methods, which permit simple animal preparation, repetitive experimental conditions, relatively medium-cost instrumentation and are performed under mild anesthesia, thus nearly under physiological conditions. A selection of recent examples of optical intrinsic imaging, fluorescence imaging and bioluminescence imaging to characterize the stroke pathology and engrafted stem cells are discussed. Special attention is paid to novel optimal reporter genes/probes for genetic labeling and tracking of stem cells and appropriate transgenic animal models. Requirements, advantages and limitations of these imaging platforms are critically discussed and placed into the context of other non-invasive techniques, e.g., magnetic resonance imaging and positron emission tomography, which can be joined with optical imaging in multimodal approaches.

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Fluorescent probes as a tool for cell population tracking in spontaneously active neural networks derived from human pluripotent stem cells

Cited by 10 publications

References 22 publications

Biological and medical applications of a brain-on-a-chip

Biological and medical applications of a brain-on-a-chip

In vitro field potential monitoring on a multi-microelectrode array for the electrophysiological long-term screening of neural stem cell maturation

A review of novel optical imaging strategies of the stroke pathology and stem cell therapy in stroke

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