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Modeling Acute Myeloid Leukemia Using StarPEG-Heparin Hydrogels
Biomimetic semi-synthetic hydrogels formed from a combination of star-shaped poly(ethylene glycol) (starPEG) and the glycosaminoglycan, heparin,...
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Organotypic 3D Cell Culture of the Embryonic Lacrimal Gland
Ectodermal organ development, including lacrimal gland, is characterized by an interaction between an epithelium and a mesenchyme. Murine lacrimal...
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Human Primordial Germ Cell-Like Cell Induction from Pluripotent Stem Cells by SOX17 and PRDM1 Expression
Human primordial germ cell (PGC) development initiates about 2 weeks after fertilization during embryogenesis. Unique molecular events follow,...
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Manufacturing of a Human Adipose-Derived Hydrogel
Hydrogels are considered a viable in vitro alternative to monolayer cultures. They provide quintessential characteristics for in vitro studies...
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A Method for Screening a Kinase Inhibitor Drug Library on ASCs
Breast cancer is an ongoing issue due to its high mortality rates. Obesity enhances the problems associated with breast cancer, meaning there must be...
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Purification, Reconstitution, and Functional Analysis of Connexin Hemichannels
Connexins are the proteins that form the gap junction channels that are essential for cell-to-cell communication. These channels are formed by...
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Spatial and Temporal Localization of Connexins in Cells Using Confocal Microscopy
The 21-member connexin family found in humans is the building block of both single-membrane spanning channels (hemichannels) and double-membrane...
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Analyzing Fcγ-Receptor Interactions on Monocytes with the Proximity Ligation Assay (PLA)
Proximity ligation assays (PLA) enable the detection and characterization of protein interactions independent of protein abundance or genetic...
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Molecular Dynamics and Other HPC Simulations for Drug Discovery
High performance computing (HPC) is taking an increasingly important place in drug discovery. It makes possible the simulation of complex biochemical...
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Bioorthogonal Labeling and Click-Chemistry-Based Visualization of the Tannerella forsythia Cell Wall
The objective of this chapter is to provide a detailed protocol for the peptidoglycan (cell wall) labeling of the periodontal pathogen Tannerella...
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Isolation of Human Villous Cytotrophoblastic Cells from Term Placenta for Transfection
Isolation of villous cytotrophoblastic cells (vCTBs) from placental tissue provides the possibility of studying differentiation of these cells as...
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Combination of CRISPR-Cas9-RNP and Single-Cell RNAseq to Identify Cell State-Specific FOXJ1 Functions in the Human Airway Epithelium
The study of the airway epithelium in vitro is routinely performed using air-liquid culture (ALI) models from nasal or bronchial basal cells. These...
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Airway Cells 3D Reconstruction via Manual and Machine-Learning Aided Segmentation of Volume EM Datasets
Volume electron microscopy (vEM) is a high-resolution imaging technique capable of revealing the 3D structure of cells, tissues, and model organisms....
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Endogenous Tagging of Ciliary Genes in Human RPE1 Cells for Live-Cell Imaging
CRISPR-mediated endogenous tagging of genes provides unique possibilities to explore the function and dynamic subcellular localization of proteins in...
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Protocols for Myosin and Actin-Myosin Assays Using Rapid, Stopped-Flow Kinetics
Fast transient kinetics using stopped-flow fluorimetry is now a powerful method for defining the ATPaseATPase cycle of myosinMyosins and its...
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Artificial Modulation and Rewiring of Cell Cycle Progression Using Synthetic Circuits in Fission Yeast
Cell cycle control is a central aspect of the biology of proliferating eukaryotic cells. However, progression through the cell cycle relies on a...
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Purification of Borrelia burgdorferi Outer Membrane Vesicles
Bacterial outer membrane vesicles (OMVs) are spherical membrane constructs shed by gram-negative bacteria. OMVs produced by the Lyme disease pathogen...
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Studying Translesion DNA Synthesis Using Xenopus In Vitro Systems
Cell-free extracts derived from Xenopus eggs have been widely used to decipher molecular pathways involved in several cellular processes including...
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CYTOSPOT: Intracellular Staining and Detection of Cytokines by Flow Cytometer, an Alternative to LYMESPOT
Intracellular cytokine staining is a versatile technique used to analyze cytokine production in individual cells by flow cytometry. This methodology...
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Systems Biology Approaches to Understanding COVID-19 Spread in the Population
In essence, the COVID-19 pandemic can be regarded as a systems biology problem, with the entire world as the system, and the human population as the...