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The Icahn School of Medicine at Mount Sinai

6 ARTICLES PUBLISHED IN JoVE

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Developmental Biology

Methods to Examine the Lymph Gland and Hemocytes in Drosophila Larvae
Theresa A. Reimels 1,2, Cathie M. Pfleger 1,2
1The Graduate School of Biomedical Sciences, The Icahn School of Medicine at Mount Sinai, 2Department of Oncological Sciences, The Icahn School of Medicine at Mount Sinai

Drosophila and mammalian hematopoietic systems share many common features, making Drosophila an attractive genetic model to study hematopoiesis. Here we demonstrate dissection and mounting of the major larval hematopoietic organ for immunohistochemistry. We also describe methods to assay various larval hematopoietic compartments including circulating hemocytes and sessile crystal cells.

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Immunology and Infection

Electrochemiluminescence Assays for Human Islet Autoantibodies
Yong Gu 1,2, Zhiyuan Zhao 1, Dongmei Miao 1, Hilary High 1, Tao Yang 2, Liping Yu 1
1Barbara Davis Center for Childhood Diabetes, University of Colorado Denver, 2Department of Endocrinology, First Affiliated Hospital of Nanjing Medical University

Here, we presented the methods to detect human islet autoantibodies using electrochemiluminescence (ECL) assays. The protocol, used to predict type 1 diabetes, can be expanded upon to detect autoantibodies for other autoimmune diseases.

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Medicine

A High-Throughput Electrochemiluminescence 7-Plex Assay Simultaneously Screening for Type 1 Diabetes and Multiple Autoimmune Diseases
Xiaofan Jia *1,2, Ling He *1,3, Yong Gu 1, Hilary High 1, Liping Yu 1
1Barbara Davis Center for Diabetes, University of Colorado School of Medicine, 2Department of Endocrinology, Beijing Hospital, National Center of Gerontology; Institute of Geriatric Medicine, Chinese Academy of Medical Sciences, 3Department of endocrinology, Guangzhou First People's Hospital, School of Medicine, South China University of Technology

We model a simple multiplexed ECL assay that combines 7 autoantibody assays together. The assay is capable of screening for T1D and multiple other autoimmune diseases, simultaneously, including celiac disease, autoimmune thyroid disease, and autoimmune polyglandular syndrome 1.

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Biology

Isolation of Mouse Interstitial Valve Cells to Study the Calcification of the Aortic Valve In Vitro
Rihab Bouchareb 1, Djamel Lebeche 1,2,3
1Cardiovascular Research Center, The Icahn School of Medicine at Mount Sinai, 2Diabetes, Obesity and Metabolism Institute, Department of Medicine, The Icahn School of Medicine at Mount Sinai, 3Graduate School of Biomedical Sciences, The Icahn School of Medicine at Mount Sinai

This article describes the isolation of mouse aortic valve cells by a two-step collagenase procedure. Isolated mouse valve cells are important for performing different assays, such as this in vitro calcification assay, and for investigating the molecular pathways leading to aortic valve mineralization.

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Immunology and Infection

A High-Throughput Multiplexed Screening for Type 1 Diabetes, Celiac Diseases, and COVID-19
Xiaofan Jia *1, Ling He *1,2, Dongmei Miao 1, Caiguo Zhang 1, Marian Rewers 1, Liping Yu 1
1Barbara Davis Center for Diabetes, University of Colorado School of Medicine, 2Department of Endocrinology, Guangzhou First People’s Hospital, School of Medicine, South China University of Technology

In line with the urgent need for screening for type 1 diabetes, celiac disease, and coronavirus disease 2019, we developed a high throughput 6-Plex electrochemiluminescence assay to simultaneously detect all four islet autoantibodies, tissue transglutaminase autoantibodies, and antibodies to the receptor binding domain of severe acute respiratory syndrome coronavirus 2.

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Biology

Isolation of Conditionally Immortalized Mouse Glomerular Endothelial Cells with Fluorescent Mitochondria
Rihab Bouchareb 1, Liping Yu 1, Emelie Lassen 1, Ilse S. Daehn 1
1Department of Medicine, Division of Nephrology, The Icahn School of Medicine at Mount Sinai

The article describes the method for isolating conditionally immortalized glomerular endothelial cells from the kidneys of transgenic mice expressing the thermolabile simian virus 40 and photo-activatable mitochondria, PhAMexcised. We describe the procedure for glomeruli isolation from whole kidneys using beads, digestion steps, seeding, and culturing of GECs-CD31 positive.

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