scholarly journals Asymmetric Arp2/3-mediated actin assembly facilitates clathrin-mediated endocytosis at stalled sites in genome-edited human stem cells.

2021 ◽  
Author(s):  
Meiyan Jin ◽  
Cyna Shirazinejad ◽  
Bowen Wang ◽  
Amy Yan ◽  
Johannes Schöneberg ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mammalian Cells ◽  
Three Dimensional ◽  
Super Resolution ◽  
Vesicle Formation ◽  
Actin Assembly ◽  
Human Stem Cells ◽  
Forming Process ◽  
Filament Assembly ◽  
Induced Pluripotent

Actin filament assembly facilitates vesicle formation in several trafficking pathways including clathrin-mediated endocytosis (CME). However, how actin assembly forces are harnessed has not been fully revealed for any vesicle forming process. In this study, three-dimensional (3D) super-resolution microscopy, live-cell imaging of triple-genome-edited, induced pluripotent stem cells (iPSCs), and newly developed machine-learning-based computational analysis tools, were used to comprehensively analyze assembly dynamics and geometry of proteins representing three different CME functional modules. When hundreds of CME events with and without associated Arp2/3-dependent actin network assembly were compared, sites with actin assembly showed a distinct delay between completion of endocytic coat expansion and vesicle scission, consistent with the notion that these were stalled sites requiring actin assembly forces to complete vesicle formation. Importantly, our analysis showed that N-WASP is preferentially recruited to one side of CME sites, where it stimulates actin assembly asymmetrically. These results indicate that in mammalian cells actin assembly is induced at stalled CME sites, where asymmetric forces pull the plasma membrane into the cell much like a bottle opener pulls off a bottle cap.

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

All About Cells

Bioprinting ◽  
2021 ◽  
pp. 21-39
Author(s):  
Kenneth Douglas
Keyword(s):  
Stem Cells ◽  
Cell Membrane ◽  
Three Dimensional ◽  
Scuba Diving ◽  
Cell Types ◽  
Two Dimensional ◽  
Cell Organelles ◽  
Outer Skin ◽  
Induced Pluripotent ◽  
Selective Access

Abstract: This chapter takes the reader on an imaginary scuba diving tour of the watery world of the cell and its surroundings, pointing out features such as the cytoskeleton (that forms the equivalent of the bones and muscles of our cells), the cell membrane (the outer skin of the cell), and the cell membrane’s embedded proteins that provide selective access to the interior of the cell—organelles (elfin versions our own organs). The chapter stresses the tumultuous action that occurs non-stop within the cells as proteins are assembled for use within and outside the cells. The chapter discusses stem cells, including the discovery of induced pluripotent stem cells. The chapter relates how cells differentiate to become dissimilar cell types, stresses the importance of three-dimensional study of cells (rather than two-dimensional study), and explains the different ways in which cells talk to each other.

scholarly journals Modeling Herpes Simplex Virus 1 Infections in Human Central Nervous System Neuronal Cells Using Two- and Three-Dimensional Cultures Derived from Induced Pluripotent Stem Cells

2019 ◽  
Vol 93 (9) ◽  
Author(s):  
Leonardo D’Aiuto ◽  
David C. Bloom ◽  
Jennifer N. Naciri ◽  
Adam Smith ◽  
Terri G. Edwards ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Three Dimensional ◽  
Two Dimensional ◽  
Herpes Simplex Virus 1 ◽  
Cns Neurons ◽  
Induced Pluripotent ◽  
Simplex Virus ◽  
Hsv 1

ABSTRACTHerpes simplex virus 1 (HSV-1) establishes latency in both peripheral nerve ganglia and the central nervous system (CNS). The outcomes of acute and latent infections in these different anatomic sites appear to be distinct. It is becoming clear that many of the existing culture models using animal primary neurons to investigate HSV-1 infection of the CNS are limited and not ideal, and most do not recapitulate features of CNS neurons. Human induced pluripotent stem cells (hiPSCs) and neurons derived from them are documented as tools to study aspects of neuropathogenesis, but few have focused on modeling infections of the CNS. Here, we characterize functional two-dimensional (2D) CNS-like neuron cultures and three-dimensional (3D) brain organoids made from hiPSCs to model HSV-1–human–CNS interactions. Our results show that (i) hiPSC-derived CNS neurons are permissive for HSV-1 infection; (ii) a quiescent state exhibiting key landmarks of HSV-1 latency described in animal models can be established in hiPSC-derived CNS neurons; (iii) the complex laminar structure of the organoids can be efficiently infected with HSV, with virus being transported from the periphery to the central layers of the organoid; and (iv) the organoids support reactivation of HSV-1, albeit less efficiently than 2D cultures. Collectively, our results indicate that hiPSC-derived neuronal platforms, especially 3D organoids, offer an extraordinary opportunity for modeling the interaction of HSV-1 with the complex cellular and architectural structure of the human CNS.IMPORTANCEThis study employed human induced pluripotent stem cells (hiPSCs) to model acute and latent HSV-1 infections in two-dimensional (2D) and three-dimensional (3D) CNS neuronal cultures. We successfully established acute HSV-1 infections and infections showing features of latency. HSV-1 infection of the 3D organoids was able to spread from the outer surface of the organoid and was transported to the interior lamina, providing a model to study HSV-1 trafficking through complex neuronal tissue structures. HSV-1 could be reactivated in both culture systems; though, in contrast to 2D cultures, it appeared to be more difficult to reactivate HSV-1 in 3D cultures, potentially paralleling the low efficiency of HSV-1 reactivation in the CNS of animal models. The reactivation events were accompanied by dramatic neuronal morphological changes and cell-cell fusion. Together, our results provide substantive evidence of the suitability of hiPSC-based neuronal platforms to model HSV-1–CNS interactions in a human context.

scholarly journals Hepatic Differentiation of Human Induced Pluripotent Stem Cells in a Perfused Three-Dimensional Multicompartment Bioreactor

2016 ◽  
Vol 5 (1) ◽  
pp. 235-248 ◽  
Author(s):  
Nora Freyer ◽  
Fanny Knöspel ◽  
Nadja Strahl ◽  
Leila Amini ◽  
Petra Schrade ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Three Dimensional ◽  
Hepatic Differentiation ◽  
Induced Pluripotent

Three-dimensional functional human myocardial tissues fabricated from induced pluripotent stem cells

2015 ◽  
Vol 11 (3) ◽  
pp. 926-935 ◽  
Author(s):  
Hyoe Komae ◽  
Hidekazu Sekine ◽  
Izumi Dobashi ◽  
Katsuhisa Matsuura ◽  
Minoru Ono ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Three Dimensional ◽  
Induced Pluripotent

Assessment of Induced Pluripotent Stem Cell-Derived Cardiomyocyte Contractility Using Micropost Arrays

2013 ◽  
Author(s):  
Marita L. Rodriguez ◽  
Charles E. Murry ◽  
Nathan J. Sniadecki
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
Heart Tissue ◽  
Patient Specific ◽  
Human Stem Cells ◽  
Cell Therapies ◽  
Cardiomyocyte Contractility ◽  
Induced Pluripotent ◽  
Contractile Forces

Cardiovascular stem cell therapies have shown increasing promise as a potential therapeutic means for reversing the effects of a myocardial infarction [1]. Out of the currently available sources of human stem cells, human induced pluripotent stem cells (hiPSCs) are very promising in that: the number of cell lines that can be induced to the pluripotent state is extremely vast, they serve as a potential source for patient-specific cardiomyocytes, and their use is non-controversial. However, before they can be used feasibly in a clinical setting, the functional engraftment of these cells into the host tissue must be improved [2]. It is hypothesized that the structural and functional maturity of the stem-cell derived cardiomyocytes prior to implantation, may significantly affect the ability of these cells to engraft with resident heart tissue [3]. One of the most important functional characteristics of a cardiomyocyte is its ability to produce contractile forces. However, assessing the contractile properties of single iPS-CMs is a difficult task. iPS-CMs generally have relatively unorganized cytoskeletons, with stress fibers in multiple directions. This trait renders one or two-point force assays ineffectual in determining total cell forces. Furthermore, iPS-CMs don’t spread well on tissue culture surfaces, which make two-dimensional force measurements almost impossible.

scholarly journals Apolipoprotein E (APOE) expression pattern in human induced pluripotent stem cells during in vitro neural induction.

2020 ◽  
Author(s):  
Hyunah Lee ◽  
Graham D. Cocks ◽  
Paulina Nowosiad ◽  
Lucia M. Dutan Polit ◽  
Jack Price ◽  
...  
Keyword(s):  
Stem Cells ◽  
Apolipoprotein E ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Neural Induction ◽  
Human Stem Cells ◽  
Peripheral Tissues ◽  
Induced Pluripotent ◽  
Apoe Expression

Abstract Apolipoprotein E (ApoE) is a multifunctional protein that plays significant roles in important cellular mechanisms in peripheral tissues and is as well expressed in the central nervous system, notably by adult neural stem cells (NSCs) in the hippocampus. Evidence from animal studies suggest that ApoE is critical for adult NSC maintenance. However, whether ApoE has the potential to play a similar role in human NSCs has not been directly investigated. To address this question, we conducted a focused study on APOE gene expression level using an in vitro model of neural differentiation and human induced pluripotent stem cells derived from a neurotypical individual. We found that APOE expression was dramatically decreased as the cells became more differentiated, indicating that APOE expression levels reflect the degree of cellular differentiation during neural induction suggesting a potential role for ApoE in human NSC maintenance. Our findings justify further investigations being carried out to understand whether changes in APOE level can directly impact the neurogenic capacity of human stem cells.

scholarly journals Intestinal Organoids—Current and Future Applications

2016 ◽  
Author(s):  
Andre M. C. Meneses ◽  
Kerstin Schneeberger ◽  
Hedwig S. Kruitwagen ◽  
Louis C. Penning ◽  
Frank G. van Steenbeek ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Pluripotent Stem Cells ◽  
Disease Modeling ◽  
Three Dimensional ◽  
Intestinal Stem Cells ◽  
Complex Structures ◽  
Technical Advances ◽  
Induced Pluripotent

Recent technical advances in the stem cell field have enabled the in vitro generation of complex structures resembling whole organs termed organoids. Most of these approaches employ culture systems that allow stem cell-derived or tissue progenitor cells to self-organize into three-dimensional (3D)-structures. Since organoids can be grown from various species, organs and from patient-derived induced pluripotent stem cells, they create significant prospects for modelling development and diseases, for toxicology and drug discovery studies, and in the field of regenerative medicine. Here, we report on intestinal stem cells, organoid culture, organoid disease modeling, transplantation, current and future uses of this exciting new insight model to veterinary medicine field.

Micro-CT Technique for Three-Dimensional Visualization of Human Stem Cells

2013 ◽  
pp. 143-152 ◽  
Author(s):  
Andrea Farini ◽  
Chiara Villa ◽  
Marzia Belicchi ◽  
Mirella Meregalli ◽  
Yvan Torrente
Keyword(s):  
Stem Cells ◽  
Three Dimensional ◽  
Micro Ct ◽  
Human Stem Cells ◽  
Ct Technique
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