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https://www.readbyqxmd.com/read/27906170/generating-high-purity-cardiac-and-endothelial-derivatives-from-patterned-mesoderm-using-human-pluripotent-stem-cells
#1
Nathan J Palpant, Lil Pabon, Clayton E Friedman, Meredith Roberts, Brandon Hadland, Rebecca J Zaunbrecher, Irwin Bernstein, Ying Zheng, Charles E Murry
Human pluripotent stem cells (hPSCs) provide a valuable model for the study of human development and a means to generate a scalable source of cells for therapeutic applications. This protocol specifies cell fate efficiently into cardiac and endothelial lineages from hPSCs. The protocol takes 2 weeks to complete and requires experience in hPSC culture and differentiation techniques. Building on lessons taken from early development, this monolayer-directed differentiation protocol uses different concentrations of activin A and bone morphogenetic protein 4 (BMP4) to polarize cells into mesodermal subtypes that reflect mid-primitive-streak cardiogenic mesoderm and posterior-primitive-streak hemogenic mesoderm...
January 2017: Nature Protocols
https://www.readbyqxmd.com/read/27830548/fetal-liver-stem-progenitor-cell-transplantation-a-model-to-study-tissue-mass-replacement-and-cell-based-therapies
#2
Mladen I Yovchev, Michael Oertel
Liver transplantation is the only therapeutic treatment for patients with end-stage liver diseases. However, donor organ scarcity is the major limitation, and therefore, alternative strategies are urgently needed. The ultimate goal for successful cell-based therapies is the ability of transplanted cells to efficiently engraft and reconstitute injured liver mass. To evaluate the repopulation capacity of transplanted cells, it is essential to identify their specific characteristics, as well as to study the mechanism(s) Through which transplanted donor cells replace tissue mass in hepatic microenvironments, using well-established cell transplantation models...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27820711/when-protocols-become-fairy-tales-and-gods-remain-buried-under-excerpts-from-the-diary-of-forensic-experts-at-ground-zero-during-the-mega-quake-that-hit-nepal
#3
Jenash Acharya, Rijen Shrestha, Pramod Kumar Shrestha, Tanuj Kanchan, Kewal Krishan
More than a year has passed since the mega quake hit Nepal. Although a lot is written about the lacunae in disaster preparedness, lessons learnt, and public health concerns after the aftermath, nothing much has been written about the fate of the deceased. Although saving takes priority after a disaster, the management of dead bodies also requires immediate attention because the process of identification becomes more difficult and expensive as time passes. This article shares the firsthand experiences of forensic experts at ground zero during the quake that may be useful to forensic experts responsible for handling such situations worldwide...
November 4, 2016: American Journal of Forensic Medicine and Pathology
https://www.readbyqxmd.com/read/27815897/synchronization-and-desynchronization-of-cells-by-interventions-on-the-spindle-assembly-checkpoint
#4
Mohamed Jemaà, Gwenola Manic, Ilio Vitale
Cell cycle checkpoints are surveillance mechanisms that sequentially and continuously monitor cell cycle progression thereby contributing to the preservation of genetic stability. Among them, the spindle assembly checkpoint (SAC) prevents the occurrence of abnormal divisions by halting the metaphase to anaphase transition following the detection of erroneous microtubules-kinetochore attachment(s). Most synchronization strategies are based on the activation of cell cycle checkpoints to enrich the population of cells in a specific phase of the cell cycle...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27807896/transient-cnp-expression-by-early-progenitors-causes-cre-lox-based-reporter-lines-to-map-profoundly-different-fates
#5
Reshmi Tognatta, Wenjing Sun, Sandra Goebbels, Klaus-Armin Nave, Akiko Nishiyama, Susanne Schoch, Leda Dimou, Dirk Dietrich
NG2 expressing oligodendroglial precursor cells are ubiquitous in the central nervous system and the only cell type cycling throughout life. Previous fate mapping studies have remained inconsistent regarding the question whether NG2 cells are capable of generating certain types of neurons. Here, we use CNP-Cre mice to map the fate of a sub-population of NG2 cells assumed to be close to differentiation. When crossing these mice with the ROSA26/YFP Cre-reporter line we discovered large numbers of reporter-expressing pyramidal neurons in the piriform and dorsal cortex...
November 3, 2016: Glia
https://www.readbyqxmd.com/read/27805614/detection-of-the-ph-dependent-activity-of-escherichia-coli-chaperone-hdeb-in-vitro-and-in-vivo
#6
Jan-Ulrik Dahl, Philipp Koldewey, James C A Bardwell, Ursula Jakob
Bacteria are frequently exposed to environmental changes, such as alterations in pH, temperature, redox status, light exposure or mechanical force. Many of these conditions cause protein unfolding in the cell and have detrimental impact on the survival of the organism. A group of unrelated, stress-specific molecular chaperones have been shown to play essential roles in the survival of these stress conditions. While fully folded and chaperone-inactive before stress, these proteins rapidly unfold and become chaperone-active under specific stress conditions...
October 23, 2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/27796959/techniques-for-the-reprogramming-of-exogenous-stem-progenitor-cell-populations-towards-a-mammary-epithelial-cell-fate
#7
Gilbert H Smith, Corinne A Boulanger
This chapter considers the techniques necessary and required for the reprogramming of exogenous stem/progenitor cell populations towards a mammary epithelial cell fate. The protocols describe how to isolate cells from alternate mouse organs such as testicles of male mice and mix them with mammary cells to generate chimeric glands comprised of male and female epithelial cells that are fully competent. During the reformation of mammary stem cell niches by dispersed epithelial cells, in the context of the intact epithelium-free mammary stroma, non-mammary cells are sequestered and reprogrammed to perform mammary epithelial cell functions including those ascribed to mammary stem/progenitor cells...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27782412/communication-thermal-unimolecular-decomposition-of-syn-ch3choo-a-kinetic-study
#8
Thanh Lam Nguyen, Laura McCaslin, Michael C McCarthy, John F Stanton
The thermal decomposition of syn-ethanal-oxide (syn-CH3CHOO) through vinyl hydrogen peroxide (VHP) leading to hydroxyl radical is characterized using a modification of the HEAT thermochemical protocol. The isomerization step of syn-CH3CHOO to VHP via a 1,4 H-shift, which involves a moderate barrier of 72 kJ/mol, is found to be rate determining. A two-dimensional master equation approach, in combination with semi-classical transition state theory, is employed to calculate the time evolution of various species as well as to obtain phenomenological rate coefficients...
October 7, 2016: Journal of Chemical Physics
https://www.readbyqxmd.com/read/27744003/neural-organoids-for-disease-phenotyping-drug-screening-and-developmental-biology-studies
#9
Brigham J Hartley, Kristen J Brennand
Human induced pluripotent stem cells (hiPSCs) can theoretically yield limitless supplies of cells fated to any cell type that comprise the human organism, making them a new tool by which to potentially overcome caveats in current biomedical research. In vitro derivation of central nervous system (CNS) cell types has the potential to provide material for drug discovery and validation, safety and toxicity assays, cell replacement therapy and the elucidation of previously unknown disease mechanisms. However, current two-dimensional (2D) CNS differentiation protocols do not faithfully recapitulate the spatial organization of heterogeneous tissue, nor the cell-cell interactions, cell-extracellular matrix interactions, or specific physiological functions generated within complex tissue such as the brain...
October 12, 2016: Neurochemistry International
https://www.readbyqxmd.com/read/27737438/assessment-of-scaffolding-properties-for-chondrogenic-differentiation-of-adipose-derived-mesenchymal-stem-cells-in-nasal-reconstruction
#10
Serban San-Marina, Ayushman Sharma, Stephen G Voss, Jeffrey R Janus, Grant S Hamilton
Importance: Nasal reconstruction in patients who are missing a significant amount of structural nasal support remains a difficult challenge. One challenge is the deficiency of cartilage left within the nose as a consequence of rhinectomy or a midline destructive disease. Historically, the standard donor source for large quantities of native cartilage has been costal cartilage. Objective: To enable the development of protocols for new mesenchymal stem cell technologies as alternative procedures with reduced donor site morbidity, risk of infection and extrusion...
October 13, 2016: JAMA Facial Plastic Surgery
https://www.readbyqxmd.com/read/27735934/self-organization-of-human-embryonic-stem-cells-on-micropatterns
#11
Alessia Deglincerti, Fred Etoc, M Cecilia Guerra, Iain Martyn, Jakob Metzger, Albert Ruzo, Mijo Simunovic, Anna Yoney, Ali H Brivanlou, Eric Siggia, Aryeh Warmflash
Fate allocation in the gastrulating embryo is spatially organized as cells differentiate into specialized cell types depending on their positions with respect to the body axes. There is a need for in vitro protocols that allow the study of spatial organization associated with this developmental transition. Although embryoid bodies and organoids can exhibit some spatial organization of differentiated cells, methods that generate embryoid bodies or organoids do not yield consistent and fully reproducible results...
November 2016: Nature Protocols
https://www.readbyqxmd.com/read/27734355/transplantation-as-a-quantitative-assay-to-study-mammalian-male-germline-stem-cells
#12
Aileen R Helsel, Jon M Oatley
In mammals, the activities of spermatogonial stem cells (SSCs) provide the foundation for continual spermatogenesis throughout a male's reproductive lifetime. At present, the defining characteristics of SSCs and mechanisms controlling their fate decisions are not well understood. Transplantation is a definitive functional measure of stem cell capacity for male germ cells that can be used as an assay to provide an unequivocal quantification of the SSC content in an experimental cell population. Here, we discuss the procedure for mice and provide protocols for preparing donor germ cell suspensions from testes directly or primary cultures of spermatogonia for transplantation, enriching for SSCs, preparing recipient males, microinjection into recipient testes, and considerations for experimental design...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27725269/size-of-monodispersed-nanomaterials-evaluated-by-dynamic-light-scattering-protocol-validated-for-measurements-of-60-and-203nm-diameter-nanomaterials-is-now-extended-to-100-and-400nm
#13
F Varenne, J Botton, C Merlet, H Hillaireau, F-X Legrand, G Barratt, C Vauthier
In vivo fate of nanomaterials is influenced by the particle size among other parameters. Thus, Health Agencies have identified the size of nanomaterial as an essential physicochemical property to characterize. This parameter can be explored by dynamic light scattering (DLS) that is described in the ISO standard 22412:2008(E) and is one of the methods recognized by Health Agencies. However, no protocol of DLS size measurement has been validated over a large range of size so far. In this work, we propose an extension of validation of a protocol of size measurement by DLS previously validated with certified reference materials (CRM) at 60 and 203nm...
October 7, 2016: International Journal of Pharmaceutics
https://www.readbyqxmd.com/read/27693946/near-infrared-light-triggered-release-of-small-molecules-for-controlled-differentiation-and-long-term-tracking-of-stem-cells-in%C3%A2-vivo-using-upconversion-nanoparticles
#14
Jinming Li, Wayne Yuk-Wai Lee, Tianyi Wu, Jianbin Xu, Kunyu Zhang, Dexter Siu Hong Wong, Rui Li, Gang Li, Liming Bian
Human mesenchymal stem cells (hMSCs) hold considerable potential for regenerative medicine, but their application is limited by the lack of an efficient method to control differentiation and track the migration of implanted cells in vivo. In this study, we developed a multifunctional nanocarrier based on upconversion nanoparticles (UCNPs) for controlling differentiation and long-term tracking of hMSCs. The UCNPs are conjugated with the peptide (Cys-Arg-Gly-Asp, CRGD) and the differentiation-inducing kartogenin (KGN) via a photocaged linker on the surface, and the obtained UCNP nanocarrier can be efficiently uptaken by hMSCs...
December 2016: Biomaterials
https://www.readbyqxmd.com/read/27670748/tracking-the-fate-of-surface-plasmon-resonance-generated-hot-electrons-by-in-situ-sers-surveying-of-catalyzed-reaction
#15
Rui Liu, Zuoliang He, Jiefang Sun, Jingfu Liu, Guibin Jiang
Plasmonic catalysis is an emerging process that utilizes surface plasmon resonance (SPR) process to harnesses solar energy for the promotion of catalyzed reactions. In most cases, SPR generated hot electrons (HEs) play an indispensable role in this solar-chemical energy shift process. Therefore, understanding the effectiveness of the HEs in promoting chemical reactions, and identifying the key factors that contribute to this utilization efficiency is of profound importance. Herein, the authors outline an in situ surface enhanced Raman spectroscopy protocol to track the fate of HEs...
September 27, 2016: Small
https://www.readbyqxmd.com/read/27668322/the-variability-of-standard-artificial-soils-cadmium-and-phenanthrene-sorption-measured-by-a-batch-equilibrium-method
#16
Lucie Bielská, Ivana Hovorková, Jan Kuta, Jiří Machát, Jakub Hofman
Artificial soil (AS) is used in soil ecotoxicology as a test medium or reference matrix. AS is prepared according to standard OECD/ISO protocols and components of local sources are usually used by laboratories. This may result in significant inter-laboratory variations in AS properties and, consequently, in the fate and bioavailability of tested chemicals. In order to reveal the extent and sources of variations, the batch equilibrium method was applied to measure the sorption of 2 model compounds (phenanthrene and cadmium) to 21 artificial soils from different laboratories...
September 23, 2016: Ecotoxicology and Environmental Safety
https://www.readbyqxmd.com/read/27631797/production-and-purification-of-recombinant-sumoylated-proteins-using-engineered-bacteria
#17
Frédérique Brockly, Marc Piechaczyk, Guillaume Bossis
SUMO is a ubiquitin-like protein that is covalently conjugated to numerous cellular proteins to modify their function and fate. Although large progresses have been made in the identification of SUMOylated proteins, the molecular consequences of their SUMOylation are generally unknown. This is, most often, due to the low abundance of SUMOylated proteins in the cell, usually less than 1 % of a given protein being modified at steady state. To gain insights into the role of specific SUMOylation targets, SUMO conjugation can be reconstituted in vitro using purified proteins...
2016: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27629765/human-induced-pluripotent-cell-derived-sensory-neurons-for-fate-commitment-of-bone-marrow-derived-schwann-cells-implications-for-remyelination-therapy
#18
Sa Cai, Lei Han, Qiang Ao, Ying-Shing Chan, Daisy Kwok-Yan Shum
: : Strategies that exploit induced pluripotent stem cells (iPSCs) to derive neurons have relied on cocktails of cytokines and growth factors to bias cell-signaling events in the course of fate choice. These are often costly and inefficient, involving multiple steps. In this study, we took an alternative approach and selected 5 small-molecule inhibitors of key signaling pathways in an 8-day program to induce differentiation of human iPSCs into sensory neurons, reaching ≥80% yield in terms of marker proteins...
September 14, 2016: Stem Cells Translational Medicine
https://www.readbyqxmd.com/read/27604750/glycogen-synthase-kinase-3-gsk3-inhibitor-sb216763-promotes-the-conversion-of-human-umbilical-cord-mesenchymal-stem-cells-into-neural-precursors-in-adherent-culture
#19
Liyang Gao, Mingyan Zhao, Peng Li, Junchao Kong, Zhijun Liu, Yonghua Chen, Rui Huang, Jiaqi Chu, Juanhua Quan, Rong Zeng
The ability to generate neural progenitor cells from human umbilical cord mesenchymal stem cells (hUC-MSCs) has provided an option to treat neurodegenerative diseases. To establish a method for this purpose, we characterized the early neural markers of hUC-MSCs-derived cells under different conditions. We found that neither the elimination of signals for alternative fate nor N2 supplement was sufficient to differentiate hUC-MSCs into neural precursor cells, but the GSK3 inhibitor SB216763 could promote an efficient neural commitment of hUC-MSCs...
September 7, 2016: Human Cell
https://www.readbyqxmd.com/read/27591140/preclinical-analysis-of-fetal-human-mesencephalic-neural-progenitor-cell-lines-characterization-and-safety-in-vitro-and-in-vivo
#20
Jisook Moon, Sigrid C Schwarz, Hyun-Seob Lee, Jun Mo Kang, Young-Eun Lee, Bona Kim, Mi-Young Sung, Günter Höglinger, Florian Wegner, Jin Su Kim, Hyung-Min Chung, Sung Woon Chang, Kwang Yul Cha, Kwang-Soo Kim, Johannes Schwarz
: : We have developed a good manufacturing practice for long-term cultivation of fetal human midbrain-derived neural progenitor cells. The generation of human dopaminergic neurons may serve as a tool of either restorative cell therapies or cellular models, particularly as a reference for phenotyping region-specific human neural stem cell lines such as human embryonic stem cells and human inducible pluripotent stem cells. We cultivated 3 different midbrain neural progenitor lines at 10, 12, and 14 weeks of gestation for more than a year and characterized them in great detail, as well as in comparison with Lund mesencephalic cells...
September 2, 2016: Stem Cells Translational Medicine
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