An Advanced Murine Model for Nonalcoholic Steatohepatitis in Association with Type 2 Diabetes
| Autor: | Katharina Schmidt-Bleek, Julia Sbierski-Kind, Jonas Kath, Joachim Spranger, Hans-Dieter Volk, Mathias Streitz |
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| Rok vydání: | 2019 |
| Předmět: |
Male
T cell General Chemical Engineering T-Lymphocytes Adipose tissue Inflammation digestive system General Biochemistry Genetics and Molecular Biology Antibodies Flow cytometry Insulin resistance Immune system Antigen Non-alcoholic Fatty Liver Disease medicine Animals medicine.diagnostic_test biology General Immunology and Microbiology business.industry General Neuroscience medicine.disease Mice Inbred C57BL Disease Models Animal medicine.anatomical_structure Adipose Tissue Diabetes Mellitus Type 2 Liver Immunology biology.protein medicine.symptom Antibody business |
| Zdroj: | Journal of visualized experiments : JoVE. (146) |
| ISSN: | 1940-087X |
| Popis: | Obesity is associated with chronic low-grade inflammation and insulin resistance, contributing to an increasing prevalence of chronic metabolic diseases, such as type 2 diabetes and nonalcoholic steatohepatitis (NASH). Recent research has established that pro-inflammatory immune cells infiltrate obese hypertrophic adipose tissue and liver. Given the emerging importance of immune cells in the context of metabolic homeostasis, there is a critical need to quantify and characterize their modification during the development of type 2 diabetes and NASH. However, animal models that induce pathophysiological features typical of human NASH are sparse. In this article, we provide a detailed protocol to identify immune cell subsets isolated from liver and adipose tissue in a reliable mouse model of NASH, established by housing high-fat diet (HFD) mice under non-specific pathogen-free (SPF) conditions without a barrier for at least seven weeks. We demonstrate the handling of mice in non-SPF conditions, digestion of the tissues and identification of macrophages, natural killer (NK) cells, dendritic cells, B and T cell subsets by flow cytometry. Representative flow cytometry plots from SPF HFD mice and non-SPF mice are provided. To obtain reliable and interpretable data, the use of antibodies, accurate and precise methods for tissue digestion and proper gating in flow cytometry experiments are critical elements. The intervention to restore physiological antigen exposure in mice by housing them in non-SPF conditions and unspecific exposure to microbial antigens could provide a relevant tool for investigating the link between immunological alterations, diet-induced obesity and related long term complications. |
| Databáze: | OpenAIRE |
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