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Confocal Microscopy of Reovirus Transport in Living Dorsal Root Ganglion Neurons

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Dissecting the Rat Mammary Gland: Isolation, Characterization, and Culture of Purified Mammary Epithelial Cells and Fibroblasts

Authors: Elizabeth A. Tovar, Rachael Sheridan, Curt J. Essenburg, Patrick S. Dischinger, Menusha Arumugam, Megan E. Callaghan, Carrie R. Graveel and Matthew R. Steensma, date: 11/20/2020, view: 250, Q&A: 0

[Abstract]

With the advent of CRISPR-Cas and the ability to easily modify the genome of diverse organisms, rat models are being increasingly developed to interrogate the genetic events underlying mammary development and tumorigenesis. Protocols for the isolation and characterization of mammary epithelial cell subpopulations have been thoroughly developed for

...

Isolation and CryoTEM of Phages Infecting Bacterial Wine Spoilers

Authors: Amel Chaïb, Marion Decossas, Cécile Philippe, Olivier Claisse, Olivier Lambert and Claire Le Marrec, date: 11/05/2020, view: 389, Q&A: 0

[Abstract]

With the objective to isolate phages infecting wine bacterial spoilers, we designed a method for the isolation and purification of phages infecting grape-associated bacteria. The method proved successful to isolate GC1 tectivirus infecting the acetic acid bacterium Gluconobacter cerinus. The isolated phage represents a new genus within the

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A Novel and Robust Single-cell Trapping Method on Digital Microfluidics

Authors: Jiao Zhai, Haoran Li, Ada Hang-Heng Wong, Cheng Dong, Shuhong Yi, Yanwei Jia, Pui-In Mak, Chuxia Deng and Rui Martins, date: 10/05/2020, view: 359, Q&A: 0

[Abstract] Due to cell heterogeneity, the differences among individual cells are averaged out in bulk analysis methods, especially in the analysis of primary tumor biopsy samples from patients. To deeply understand the cell-to-cell variation in a primary tumor, single-cell culture and analysis with limited amount of cells are in high demand. Microfluidics ...

Probe-Seq: Method for RNA Sequencing of Specific Cell Types from Animal Tissue

Authors: Ryoji Amamoto, Mauricio D. Garcia, Emma R. West, Jiho Choi, Sylvain W. Lapan, Elizabeth A. Lane, Norbert Perrimon and Constance L. Cepko, date: 09/20/2020, view: 901, Q&A: 0

[Abstract] Most organs and tissues are composed of many types of cells. To characterize cellular state, various transcription profiling approaches are currently available, including whole-tissue bulk RNA sequencing, single cell RNA sequencing (scRNA-Seq), and cell type-specific RNA sequencing. What is missing in this repertoire is a simple, versatile method ...

A Method to Efficiently Cryopreserve Mammalian Cells on Paper Platforms

Authors: Muhammedin Deliorman, Pavithra Sukumar, Roaa Alnemari and Mohammad A. Qasaimeh, date: 09/20/2020, view: 767, Q&A: 0

[Abstract] This protocol describes a simple method to cryopreserve mammalian cells within filter papers as an alternative to conventional slow-freezing approach. The method involves treating paper fibers with fibronectin, using low concentrations of the cryoprotectant dimethyl sulfoxide (DMSO), and slow freezing cells to -80 °C at a 1 °C min^-1 ...

Isolation and ex vivo Expansion of Human Limbal Epithelial Progenitor Cells

Authors: Naresh Polisetti, Günther Schlunck, Thomas Reinhard, Friedrich E. Kruse and Ursula Schlötzer-Schrehardt, date: 09/20/2020, view: 459, Q&A: 0

[Abstract] Limbal stem cell transplantation has been used successfully to treat patients with limbal stem cell deficiency all over the world. However, long term clinical results often proved less satisfactory due to the low quality of the graft or inadequate properties of transplanted cells. To enhance the ex vivo expansion of human limbal ...

In vitro Cultivation and Visualization of Malaria Liver Stages in Primary Simian Hepatocytes

Authors: Devendra Kumar Gupta and Thierry Diagana, date: 08/20/2020, view: 807, Q&A: 0

[Abstract] Human liver is the primary and obligatory site for malaria infection where sporozoites invade host hepatocytes. Malaria hepatic stages are asymptomatic and represent an attractive target for development of anti-malarial interventions and vaccines. However, owing to lack of robust and reproducible in vitro culture system, it is difficult ...

Co-culture of Murine Neurons Using a Microfluidic Device for The Study of Tau Misfolding Propagation

Authors: Grace I Hallinan, Dianne M Lopez, Mariana Vargas-Caballero, Jonathan West and Katrin Deinhardt, date: 08/20/2020, view: 916, Q&A: 0

[Abstract] The deposition of misfolded, aggregated tau protein is a hallmark of several neurodegenerative diseases, collectively termed “tauopathies”. Tau pathology spreads throughout the brain along connected pathways in a prion-like manner. The process of tau pathology propagation across circuits is a focus of intense research and has been investigated in ...

Measuring Cell Growth and Junction Development in Epithelial Cells Using Electric Cell-Substrate Impedance Sensing (ECIS)

Authors: Shaista Anwer and Katalin Szaszi, date: 08/20/2020, view: 1273, Q&A: 0

[Abstract] Electric Cell-substrate Impedance Sensing (ECIS) is an automated method that can be used to quantify processes such as cell attachment, growth, migration and barrier functions (i.e., the properties of tight junctions). The method provides simultaneous information on cell number and tight junction function by detecting electric parameters ...

Generation of Functional Mouse Hippocampal Neurons

Authors: Francesco Tomassoni-Ardori, Zhenyi Hong, Gianluca Fulgenzi and Lino Tessarollo, date: 08/05/2020, view: 1348, Q&A: 0

[Abstract] Primary culture of mouse hippocampal neurons is a very useful in vitro model for studying neuronal development, axonal and dendritic morphology, synaptic functions, and many other neuronal features. Here we describe a step-by-step process of generating primary neurons from mouse embryonic hippocampi (E17.5/E18.5). Hippocampal neurons ...

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