Honing the Double-Edged Sword: Improving Human iPSC-Microglia Models

Human induced Pluripotent Stem Cell (hiPSC) models are a valuable new tool for research into neurodegenerative diseases. Neuroinflammation is now recognized as a key process in neurodegenerative disease and aging, and microglia are central players in this. A plethora of hiPSC-derived microglial models have been published recently to explore neuroinflammation, ranging from monoculture through to xenotransplantation. However, combining physiological relevance, reproducibility, and scalability into one model is still a challenge. We examine key features of the in vitro microglial environment, especially media composition, extracellular matrix, and co-culture, to identify areas for improvement in current hiPSC-microglia models.

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Persistence of intramyocardially transplanted murine induced pluripotent stem cell-derived cardiomyocytes from different developmental stages

Stem Cell Research & Therapy ◽

10.1186/s13287-020-02089-5 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Gabriel Peinkofer ◽

Martina Maass ◽

Kurt Pfannkuche ◽

Agapios Sachinidis ◽

Stephan Baldus ◽

...

Keyword(s):

Extracellular Matrix ◽

Stem Cell ◽

Cell Adhesion ◽

Developmental Stage ◽

Pluripotent Stem Cell ◽

Induced Pluripotent Stem Cell ◽

Induced Pluripotent

Abstract Background Induced pluripotent stem cell-derived cardiomyocytes (iPSC-CM) are regarded as promising cell type for cardiac cell replacement therapy, but it is not known whether the developmental stage influences their persistence and functional integration in the host tissue, which are crucial for a long-term therapeutic benefit. To investigate this, we first tested the cell adhesion capability of murine iPSC-CM in vitro at three different time points during the differentiation process and then examined cell persistence and quality of electrical integration in the infarcted myocardium in vivo. Methods To test cell adhesion capabilities in vitro, iPSC-CM were seeded on fibronectin-coated cell culture dishes and decellularized ventricular extracellular matrix (ECM) scaffolds. After fixed periods of time, stably attached cells were quantified. For in vivo experiments, murine iPSC-CM expressing enhanced green fluorescent protein was injected into infarcted hearts of adult mice. After 6–7 days, viable ventricular tissue slices were prepared to enable action potential (AP) recordings in transplanted iPSC-CM and surrounding host cardiomyocytes. Afterwards, slices were lysed, and genomic DNA was prepared, which was then used for quantitative real-time PCR to evaluate grafted iPSC-CM count. Results The in vitro results indicated differences in cell adhesion capabilities between day 14, day 16, and day 18 iPSC-CM with day 14 iPSC-CM showing the largest number of attached cells on ECM scaffolds. After intramyocardial injection, day 14 iPSC-CM showed a significant higher cell count compared to day 16 iPSC-CM. AP measurements revealed no significant difference in the quality of electrical integration and only minor differences in AP properties between d14 and d16 iPSC-CM. Conclusion The results of the present study demonstrate that the developmental stage at the time of transplantation is crucial for the persistence of transplanted iPSC-CM. iPSC-CM at day 14 of differentiation showed the highest persistence after transplantation in vivo, which may be explained by a higher capability to adhere to the extracellular matrix.

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Three-dimensional induced pluripotent stem-cell models of human brain angiogenesis

10.21203/rs.2.18502/v1 ◽

2019 ◽

Author(s):

Peter Searson ◽

Raleigh M. Linville ◽

Diego Arevalo ◽

Joanna C. Maressa ◽

Nan Zhao

Keyword(s):

Extracellular Matrix ◽

Stem Cell ◽

Blood Brain Barrier ◽

Pluripotent Stem Cell ◽

Chemical Cues ◽

Three Dimensional ◽

Induced Pluripotent Stem Cell ◽

Induced Pluripotent ◽

Brain Angiogenesis

Abstract Background: During brain development, chemical cues released by developing neurons, cellular signaling with pericytes, and mechanical cues within the brain extracellular matrix (ECM) promotes angiogenesis occurs of brain microvascular endothelial cells (BMECs). During brain disease, angiogenesis can also occur due to pathological chemical, cellular, and mechanical signaling. Existing in vitro and in vivo models of brain angiogenesis have key limitations. Methods: Here, we develop a high-throughput in vitro BBB bead assay of brain angiogenesis utilizing 150 μm diameter beads coated with induced pluripotent stem-cell (iPSC)-derived human BMECs (dhBMECs). After embedding the beads within a 3D matrix, we introduce various chemical cues and extracellular matrix components to explore their effects on angiogenic behavior. Based on the results from the bead assay, we generate a multi-scale model of the human cerebrovasculature within perfusable three-dimensional tissue-engineered blood-brain barrier (BBB) microvessels. Results: A sprouting phenotype is optimized in confluent monolayers of dhBMECs using chemical treatment with vascular endothelial growth factor (VEGF) and wnt ligands, and the inclusion of pro-angiogenic ECM components. As a proof-of-principle that the bead angiogenesis assay can be applied to study pathological angiogenesis, we show that oxidative stress can exert concentration-dependent effects on angiogenesis. Finally, we demonstrate the formation of a hierarchical microvascular model of the human blood-brain barrier displaying key structural hallmarks. Conclusions: We develop two in vitro models of brain angiogenesis: the BBB bead assay and the tissue-engineered BBB microvessel model. These platforms provide a tool kit for studies of physiological and pathological brain angiogenesis, with key advantages over existing two-dimensional models.

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Differentiation, Evaluation, and Application of Human Induced Pluripotent Stem Cell–Derived Endothelial Cells

Arteriosclerosis Thrombosis and Vascular Biology ◽

10.1161/atvbaha.117.309962 ◽

2017 ◽

Vol 37 (11) ◽

pp. 2014-2025 ◽

Cited By ~ 33

Author(s):

Yang Lin ◽

Chang-Hyun Gil ◽

Mervin C. Yoder

Keyword(s):

Endothelial Cells ◽

Stem Cell ◽

Pluripotent Stem Cell ◽

Disease Modeling ◽

Induced Pluripotent Stem Cell ◽

Patient Specific ◽

Induced Pluripotent ◽

Human Ipsc

The emergence of induced pluripotent stem cell (iPSC) technology paves the way to generate large numbers of patient-specific endothelial cells (ECs) that can be potentially delivered for regenerative medicine in patients with cardiovascular disease. In the last decade, numerous protocols that differentiate EC from iPSC have been developed by many groups. In this review, we will discuss several common strategies that have been optimized for human iPSC-EC differentiation and subsequent studies that have evaluated the potential of human iPSC-EC as a cell therapy or as a tool in disease modeling. In addition, we will emphasize the importance of using in vivo vessel-forming ability and in vitro clonogenic colony–forming potential as a gold standard with which to evaluate the quality of human iPSC-EC derived from various protocols.

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Use of 3D culture systems to generate human induced pluripotent stem cell-derived [beta]-cells in vitro

Endocrine Abstracts ◽

10.1530/endoabs.57.003 ◽

2018 ◽

Author(s):

Fantuzzi Federica ◽

Toivonen Sanna ◽

Schiavo Andrea Alex ◽

Pachera Nathalie ◽

Rajaei Bahareh ◽

...

Keyword(s):

Stem Cell ◽

Beta Cells ◽

Pluripotent Stem Cell ◽

3D Culture ◽

Induced Pluripotent Stem Cell ◽

Culture Systems ◽

Induced Pluripotent

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Cues from human atrial extracellular matrix enrich the atrial differentiation of human induced pluripotent stem cell-derived cardiomyocytes

Biomaterials Science ◽

10.1039/d0bm01686a ◽

2021 ◽

Author(s):

Fernanda C. P. Mesquita ◽

Jacquelynn Morrissey ◽

Po-Feng Lee ◽

Gustavo Monnerat ◽

Yutao Xi ◽

...

Keyword(s):

Extracellular Matrix ◽

Stem Cell ◽

Pluripotent Stem Cell ◽

Induced Pluripotent Stem Cell ◽

Cardiac Differentiation ◽

Twofold Increase ◽

Decellularized Extracellular Matrix ◽

Induced Pluripotent

Decellularized extracellular matrix (dECM) from human atria preserves key native components that directed the cardiac differentiation of hiPSCs to an atrial-like phenotype, yielding a twofold increase of functional atrial-like cells.

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