Flow cytometry —Cell isolation —Cell isolation and culture —Cell Biology

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: 1157, 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

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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: 1833, 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 ...

Isolation and High Throughput Flow Cytometric Apoptosis Assay of Human Neutrophils to Enable Compound Library Screening

Authors: K. D. Herman, A. Rahman and L. R. Prince, date: 06/05/2020, view: 1475, Q&A: 0

[Abstract] The study of human neutrophils in vitro is challenging due to their short half-life and propensity for activation. However, with careful handling and manipulation in the laboratory, they can be a powerful tool to investigate immune responses in health and disease. Here we describe a method for the isolation of human neutrophils from ...

Preparation of Single Epithelial Cells Suspension from Mouse Mammary Glands

Authors: Heng Sun, Xiaoling Xu and Chuxia Deng, date: 02/20/2020, view: 1864, Q&A: 0

[Abstract] Single cell RNA sequencing is a very powerful means for cellular heterogeneous studies and so becoming wildly utilized nowadays. To guarantee the success of such analysis, it is very important, though sometimes difficult, to obtain single cells suspension with high quality, especially from the primary solid organs like mammary glands. Digestion of ...

Quantifying HIV-1-Mediated Gut CD4⁺ T Cell Death in the Lamina Propria Aggregate Culture (LPAC) Model

Authors: Stephanie M. Dillon, Kejun Guo, Moriah J. Castleman, Mario L. Santiago and Cara C. Wilson, date: 01/20/2020, view: 1627, Q&A: 0

[Abstract] Gut CD4 T cells are major targets of HIV-1 and are massively depleted early during infection. To better understand the mechanisms governing HIV-1-mediated CD4 T cell death, we developed the physiologically-relevant Lamina Propria Aggregate Culture (LPAC) model. The LPAC model is ideal for studying CD4 T cell death induced by clinically-relevant ...

Platelet Isolation and Activation Assays

Authors: Laura C. Burzynski, Nicholas Pugh and Murray C.H. Clarke, date: 10/20/2019, view: 4111, Q&A: 0

[Abstract] Platelets regulate hemostasis and are the key determinants of pathogenic thrombosis following atherosclerotic plaque rupture. Platelets circulate in an inactive state, but become activated in response to damage to the endothelium, which exposes thrombogenic material such as collagen to the blood flow. Activation results in a number of responses, ...

Isolation and Long-term Cultivation of Mouse Alveolar Macrophages

Authors: Clara Jana-Lui Busch, Jérémy Favret, Laufey Geirsdóttir, Kaaweh Molawi and Michael H. Sieweke, date: 07/20/2019, view: 5153, Q&A: 0

[Abstract] Alveolar macrophages (AM) are tissue-resident macrophages that colonize the lung around birth and can self-maintain long-term in an adult organism without contribution of monocytes. AM are located in the pulmonary alveoli and can be harvested by washing the lungs using the method of bronchoalveolar lavage (BAL). Here, we compared different ...

In vitro Infection of Primary Human Monocytes with HIV-1

Authors: Patrick O’Connell, Yong-Hui Zheng, Andrea Amalfitano and Yasser A. Aldhamen, date: 07/05/2019, view: 2738, Q&A: 0

[Abstract] Monocyte infection by HIV-1 is an important component of chronic HIV pathogenesis. Following infection by HIV-1, monocytes are able to cross the blood brain barrier and set up a viral reservoir in the central nervous system. Additionally, in the setting of chronic HIV-1 infection, monocytes can become activated either directly through HIV-1 ...

Induction and Analysis of Anti-CD40-induced Colitis in Mice

Authors: Barbara Joyce-Shaikh, Daniel J. Cua and David Bauché, date: 02/05/2019, view: 5586, Q&A: 0

[Abstract] Colon inflammation or colitis affects more than 1 million people worldwide. Several pre-clinical models, including chemical-induced (i.e., DSS, TNBS) or pathogen-induced (i.e., Citrobacter rodentium) have been used to study mechanisms involved in the development and regulation of colitis. Anti-CD40 induced colitis model ...

Collagenase-based Single Cell Isolation of Primary Murine Brain Endothelial Cells Using Flow Cytometry

Authors: Cathrin J. Czupalla, Hanadie Yousef, Tony Wyss-Coray and Eugene C. Butcher, date: 11/20/2018, view: 6546, Q&A: 0

[Abstract] The brain endothelium is a highly specialized vascular structure that maintains the activity and integrity of the central nervous system (CNS). Previous studies have reported that the integrity of the brain endothelium is compromised in a plethora of neuropathologies. Therefore, it is of particular interest to establish a method that enables ...

Cell Biology

Generation of Giant Unilamellar Vesicles (GUVs) Using Polyacrylamide Gels

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