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https://www.readbyqxmd.com/read/29155748/using-in-vitro-live-cell-imaging-to-explore-chemotherapeutics-delivered-by-lipid-based-nanoparticles
#1
Ann L B Seynhaeve, Timo L M Ten Hagen
Conventional imaging techniques can provide detailed information about cellular processes. However, this information is based on static images in an otherwise dynamic system, and successive phases are easily overlooked or misinterpreted. Live-cell imaging and time-lapse microscopy, in which living cells can be followed for hours or even days in a more or less continuous fashion, are therefore very informative. The protocol described here allows for the investigation of the fate of chemotherapeutic nanoparticles after the delivery of doxorubicin (dox) in living cells...
November 1, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29155562/quantification-of-nanomaterial-nanomedicine-trafficking-in-vivo
#2
Liming Wang, Liang Yan, Jing Liu, Chunying Chen, Yuliang Zhao
Absorption, distribution, metabolism and excretion (ADME) of nanomaterials (NMs) or nanomedicines (NMs), are fundamental biological processes that dominate their nanomedical functions or nanotoxicities. Mostly, NMs exist in biological systems as a form of nanoparticles (NPs). However, to determine the composition, spatial distribution, and states of existence of NMs/NPs in vivo, upon entry into the body, as well as to monitor the dynamic changes of these information over time remain a challenge for the current analytical methodology due to their limited efficiently in characterizing interactions between these complex(nano)-complex(bio) systems...
November 20, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/29149762/interaction-of-biologically-relevant-ions-and-organic-molecules-with-titanium-oxide-rutile-surfaces-a-review-on-molecular-dynamics-studies
#3
REVIEW
Azade YazdanYar, Ulrich Aschauer, Paul Bowen
The surface of a biomaterial can play a major role in its biological fate since the surface is the primary pathway for its interaction with the body. As the natural response of the body to a foreign material is to encapsulate it with a fibrous material, the interactions between the body and the biomaterial are mediated by this fibrous layer. Initial interactions occur between the biomaterial surface, water, ionic species and organic molecules, which then mediate further interactions with body tissues. Surface engineering can influence these interactions and hence, improve the biocompatibility of the biomaterial...
November 7, 2017: Colloids and Surfaces. B, Biointerfaces
https://www.readbyqxmd.com/read/29147115/effects-of-aminoglycoside-antibiotics-on-human-embryonic-stem-cell-viability-during-differentiation-in-vitro
#4
Divya S Varghese, Shama Parween, Mustafa T Ardah, Bright Starling Emerald, Suraiya A Ansari
Human embryonic stem cells (hESCs) are being used extensively in array of studies to understand different mechanisms such as early human embryogenesis, drug toxicity testing, disease modeling, and cell replacement therapy. The protocols for the directed differentiation of hESCs towards specific cell types often require long-term cell cultures. To avoid bacterial contamination, these protocols include addition of antibiotics such as pen-strep and gentamicin. Although aminoglycosides, streptomycin, and gentamicin have been shown to cause cytotoxicity in various animal models, the effect of these antibiotics on hESCs is not clear...
2017: Stem Cells International
https://www.readbyqxmd.com/read/29140566/mesenchymal-stem-cell-preparation-and-transfection-free-ferumoxytol-labeling-for-mri-cell-tracking
#5
Li Liu, Chien Ho
Mesenchymal stem cells (MSCs) are multipotent cells and are the most widely studied cell type for stem cell therapies. In vivo cell tracking of MSCs labeled with an FDA-approved superparamagnetic iron-oxide (SPIO) particle by magnetic resonance imaging (MRI) provides essential information, e.g., MSC engraftment, survival, and fate, thus improving cell therapy accuracy. However, current methodology for labeling MSCs with Ferumoxytol (Feraheme(®) ), the only FDA-approved SPIO particle, needs transfection agents...
November 15, 2017: Current Protocols in Stem Cell Biology
https://www.readbyqxmd.com/read/29123019/photoredox-catalyzed-deuteration-and-tritiation-of-pharmaceutical-compounds
#6
Yong Yao Loh, Kazunori Nagao, Andrew J Hoover, David Hesk, Nelo R Rivera, Steven L Colletti, Ian W Davies, David W C MacMillan
Deuterium- and tritium-labeled pharmaceutical compounds are pivotal diagnostic tools in pharmaceutical drug discovery research, providing vital information about the biological fate of drugs and drug metabolites. Herein, we demonstrate that a photoredox-mediated hydrogen atom transfer (HAT) protocol can efficiently and selectively install deuterium and tritium at α-amino sp(3) C-H bonds in a single step, using isotopically labeled water (D2O or T2O) as the source of hydrogen isotope. In this context, we also report a convenient synthesis of T2O from T2, providing access to high specific activity T2O...
November 9, 2017: Science
https://www.readbyqxmd.com/read/29082485/tools-to-measure-cell-health-and-cytotoxicity-using-high-content-imaging-and-analysis
#7
Bhaskar S Mandavilli, Robert J Aggeler, Kevin M Chambers
High content screening (HCS)-based multiparametric measurements are very useful in early toxicity testing and safety assessment during drug development, and useful in evaluating the impact from new food supplements and environmental toxicants. Mitochondrial membrane potential, plasma membrane permeability, oxidative stress, phosphoplipidosis, and steatosis are a few of the important markers routinely studied for the assessment of drug-induced liver injury and toxicity. Mitochondrial dysfunction leads to oxidative stress and cell death...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29081882/derivation-of-telencephalic-oligodendrocyte-progenitors-from-human-pluripotent-stem-cells
#8
Tamara Major, Ann Powers, Viviane Tabar
Oligodendrocytes are the main myelinating cell of the adult CNS and are vulnerable to injury in diverse disorders such as spinal cord injury, stroke, trauma, pharmacological and radiation toxicity, as well as neuroinflammation. Human pluripotent stem cells are attractive sources of oligodendrocyte lineage cells and provide a promising treatment strategy for exogenous myelin repair through transplantation. This unit describes a protocol for the step-wise differentiation of forebrain late oligodendrocyte progenitor cells (OPCs) from human pluripotent stem cells in defined chemical in vitro culture conditions...
November 8, 2017: Current Protocols in Stem Cell Biology
https://www.readbyqxmd.com/read/29080741/design-assembly-production-and-transfection-of-synthetic-modified-mrna
#9
Sanders Oh, John A Kessler
Expression of exogenous genes using synthetic modified mRNAs has a superior safety profile and higher efficiency than DNA-based techniques. However, mRNA stability and immunogenicity have been significant barriers for wide spread use. The modified mRNAs in this protocol address these shortcomings and can be used to direct cell fate specification or cellular reprogramming.
November 7, 2017: Methods: a Companion to Methods in Enzymology
https://www.readbyqxmd.com/read/29079547/epigenetic-changes-in-mesenchymal-stem-cells-differentiation
#10
REVIEW
Ibrahim Mortada, Rola Mortada
Epigenetic factors are known to play a major role in determining stem cell fate and differentiation. Mesenchymal stem cells are the most studied population of stem cells due to their important applications in experimental biology and regenerative medicine. After a brief overview on mesenchymal stem cells, this review aims to highlight the role of epigenetic changes on mesenchymal stem cells biology and differentiation protocols with a focus on osteocytic, chondrocytic and adipocytic differentiation. Chromatin remodeling, DNA methylation, histone modifications and miRNA expression will be investigated...
October 24, 2017: European Journal of Medical Genetics
https://www.readbyqxmd.com/read/29071684/immunophenotypic-identification-of-early-myeloerythroid-development
#11
Cornelis J H Pronk, David Bryder
Myeloerythroid-restricted precursor cells, derived from multipotent hematopoietic stem cells, give rise to mature cells of the granulocyte, monocyte, erythroid, and/or thrombocytic lineages. High-resolution profiling of the developmental stages, from hematopoietic stem cells to mature progeny, is important to be able to study and understand the underlying mechanisms that guide various cell fate decisions. Also, this approach opens for greater insights into pathogenic events such as leukemia, diseases that are most often characterized by halted differentiation at defined immature precursor levels...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29055001/listeria-monocytogenes-replicate-in-bone-marrow-derived-cd11c-cells-but-not-in-dendritic-cells-isolated-from-the-murine-gastrointestinal-tract
#12
Grant S Jones, Victoria C Smith, Sarah E F D'Orazio
Recent fate-mapping studies and gene-expression profiles suggest that commonly used protocols to generate bone marrow-derived cultured dendritic cells yield a heterogeneous mixture, including some CD11c(hi) cells that may not have a bona fide counterpart in vivo. In this study, we provide further evidence of the discordance between ex vivo-isolated and in vitro-cultured CD11c(+) cells by analyzing an additional phenotype, the ability to support cytosolic growth of the facultative intracellular bacterial pathogen Listeria monocytogenes Two days after foodborne infection of mice with GFP-expressing L...
October 20, 2017: Journal of Immunology: Official Journal of the American Association of Immunologists
https://www.readbyqxmd.com/read/29053691/quantitative-whole-mount-immunofluorescence-analysis-of-cardiac-progenitor-populations-in-mouse-embryos
#13
Evan Bardot, Nikos Tzavaras, Deanna L Benson, Nicole C Dubois
The use of ever-advancing imaging techniques has contributed broadly to our increased understanding of embryonic development. Pre-implantation development and organogenesis are two areas of research that have benefitted greatly from these advances, due to the high quality of data that can be obtained directly from imaging pre-implantation embryos or ex vivo organs. While pre-implantation embryos have yielded data with especially high spatial resolution, later stages have been less amenable to three-dimensional reconstruction...
October 12, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29052205/localization-of-chromatin-marks-in-arabidopsis-early-embryos
#14
Marcelina García-Aguilar, Daphné Autran
During early embryo development, profound changes in chromatin structure and regulation take place. It is difficult to study these changes in plant embryos however, largely because of their relative inaccessibility, which impedes the application of current epigenomic and biochemistry protocols. To circumvent this issue and to analyze the epigenetic status of the embryo at both the cellular and subcellular level, we describe here a simple method to immunolocalize chromatin marks in whole mount early Arabidopsis embryos, either within maternal tissues or isolated from seeds...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29051326/multimodal-imaging-of-small-hard-retinal-drusen-in-young-healthy-adults
#15
Hilde R Pedersen, Stuart J Gilson, Alfredo Dubra, Inger Christine Munch, Michael Larsen, Rigmor C Baraas
BACKGROUND: Small hard macular drusen can be observed in the retina of adults as young as 18 years of age. Here, we seek to describe the in vivo topography and geometry of these drusen. METHODS: Retinal images were acquired in young, healthy adults using colour fundus photography, spectral domain optic coherence tomography (SD-OCT), reflectance flood-illuminated adaptive optic ophthalmoscopy (AO flood) and reflectance adaptive optic scanning light ophthalmoscopy (AOSLO) in both confocal and non-confocal split-detection modalities...
October 19, 2017: British Journal of Ophthalmology
https://www.readbyqxmd.com/read/29044469/co-differentiation-of-human-pluripotent-stem-cells-derived-cardiomyocytes-and-endothelial-cells-from-cardiac-mesoderm-provides-a-three-dimensional-model-of-cardiac-microtissue
#16
Elisa Giacomelli, Milena Bellin, Valeria V Orlova, Christine L Mummery
The formation of cardiac mesodermal subtypes is highly regulated in time and space during heart development. In vitro models based on human pluripotent stem cells (hPS cells) provide opportunities to study mechanisms underlying fate choices governing lineage specification from common cardiovascular progenitors in human embryos. The generation of cardiac endothelial cells in particular allows the creation of complex models of cardiovascular disorders in which either cardiomyocytes or endothelial cells are affected...
October 18, 2017: Current Protocols in Human Genetics
https://www.readbyqxmd.com/read/29023202/transplantation-of-zebrafish-cells-by-conventional-pneumatic-microinjector
#17
Ming Shao, Xiao-Ning Cheng, Yuan-Yuan Liu, Ji-Tong Li, De-Li Shi
Generating chimeric zebrafish by transplantation is extremely useful for live imaging in developmental, stem cell, and cancer biology, and to answer the questions of how cells acquire, keep, and/or change their fate. However, as it is technically challenging, the use of transplantation approach remains very limited by the zebrafish community. In this study, we show that this cell grafting operation can be easily achieved by using a conventional pneumatic microinjector normally used for microinjections. Compared with previously published protocols, which need additional transplantation apparatus, this alternative transplantation method works well, but needs a simpler experimental setup, and is more accessible to all investigators...
October 12, 2017: Zebrafish
https://www.readbyqxmd.com/read/29020617/live-imaging-reveals-that-the-first-division-of-differentiating-human-embryonic-stem-cells-often-yields-asymmetric-fates
#18
Katharine Brown, Kyle M Loh, Roel Nusse
How do stem cells respond to signals to initiate differentiation? Here, we show that, despite uniform exposure to differentiation-inducing extracellular signals, individual human embryonic stem cells (hESCs) respond heterogeneously. To track how hESCs incipiently exit pluripotency, we established a system to differentiate hESCs as single cells and conducted live imaging to track their very first cell division. We followed the fate of their earliest daughters as they remained undifferentiated or differentiated toward the primitive streak (the earliest descendants of pluripotent cells)...
October 10, 2017: Cell Reports
https://www.readbyqxmd.com/read/28994764/patterning-bioactive-proteins-or-peptides-on-hydrogel-using-photochemistry-for-biological-applications
#19
Taylor B Dorsey, Alexander Grath, Cancan Xu, Yi Hong, Guohao Dai
There are many biological stimuli that can influence cell behavior and stem cell differentiation. General cell culture approaches rely on soluble factors within the medium to control cell behavior. However, soluble additions cannot mimic certain signaling motifs, such as matrix-bound growth factors, cell-cell signaling, and spatial biochemical cues, which are common influences on cells. Furthermore, biophysical properties of the matrix, such as substrate stiffness, play important roles in cell fate, which is not easily manipulated using conventional cell culturing practices...
September 15, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28990928/mouse-embryonic-stem-cells-can-differentiate-via-multiple-paths-to-the-same-state
#20
James Alexander Briggs, Victor C Li, Seungkyu Lee, Clifford J Woolf, Allon Klein, Marc W Kirschner
In embryonic development, cells differentiate through stereotypical sequences of intermediate states to generate particular mature fates. By contrast, driving differentiation by ectopically expressing terminal transcription factors (direct programming) can generate similar fates by alternative routes. How differentiation in direct programming relates to embryonic differentiation is unclear. We applied single-cell RNA sequencing to compare two motor neuron differentiation protocols: a standard protocol approximating the embryonic lineage, and a direct programming method...
October 9, 2017: ELife
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