PMID- 36046625 OWN - NLM STAT- MEDLINE DCOM- 20231012 LR - 20231012 IS - 2667-2375 (Electronic) IS - 2667-2375 (Linking) VI - 2 IP - 8 DP - 2022 Aug 22 TI - An improved organotypic cell culture system to study tissue-resident macrophages ex vivo. PG - 100260 LID - 10.1016/j.crmeth.2022.100260 [doi] LID - 100260 AB - Tissue-resident macrophages (TRMs) perform organ-specific functions that are dependent on factors such as hematopoietic origin, local environment, and biological influences. A diverse range of in vitro culture systems have been developed to decipher TRM functions, including bone marrow-derived macrophages (BMDMs), induced pluripotent stem cell (iPSC)-derived TRMs, or immortalized cell lines. However, despite the usefulness of such systems, there are notable limitations. Attempts to culture primary macrophages often require purification of cells and lack a high cell yield and consistent phenotype. Here, we aimed to address these limitations by establishing an organotypic primary cell culture protocol. We obtained long-term monocultures of macrophages derived from distinct organs without prior purification using specific growth factors and tissue normoxic conditions that largely conserved a TRM-like identity in vitro. Thus, this organotypic system offers an ideal screening platform for primary macrophages from different organs that can be used for a wide range of assays and readouts. CI - (c) 2022 The Author(s). FAU - Aktories, Philipp AU - Aktories P AD - Institute of Neuropathology, Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany. AD - Faculty of Biology, University of Freiburg, 79104 Freiburg, Germany. FAU - Petry, Philippe AU - Petry P AD - Institute of Neuropathology, Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany. AD - Faculty of Biology, University of Freiburg, 79104 Freiburg, Germany. FAU - Glatz, Paulo AU - Glatz P AD - Institute of Neuropathology, Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany. AD - Faculty of Biology, University of Freiburg, 79104 Freiburg, Germany. FAU - Andrieux, Geoffroy AU - Andrieux G AD - Institute of Medical Bioinformatics and Systems Medicine, Medical Center, Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany. FAU - Oschwald, Alexander AU - Oschwald A AD - Institute of Neuropathology, Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany. AD - Faculty of Biology, University of Freiburg, 79104 Freiburg, Germany. FAU - Botterer, Hannah AU - Botterer H AD - Institute of Neuropathology, Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany. AD - Faculty of Biology, University of Freiburg, 79104 Freiburg, Germany. FAU - Gorka, Oliver AU - Gorka O AD - Institute of Neuropathology, Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany. FAU - Erny, Daniel AU - Erny D AD - Institute of Neuropathology, Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany. AD - Berta-Ottenstein-Program for Advanced Clinician Scientists, Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany. FAU - Boerries, Melanie AU - Boerries M AD - Institute of Medical Bioinformatics and Systems Medicine, Medical Center, Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany. AD - German Cancer Consortium (DKTK), Partner Site Freiburg, and German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany. FAU - Henneke, Philipp AU - Henneke P AD - CIBSS-Center for Integrative Biological Signaling Studies, University of Freiburg, 79104 Freiburg, Germany. AD - Institute for Immunodeficiency, Center for Chronic Immunodeficiency, and Center for Pediatrics and Adolescent Medicine, Medical Center, Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany. FAU - Gross, Olaf AU - Gross O AD - Institute of Neuropathology, Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany. AD - Center for Basics in NeuroModulation (NeuroModulBasics), Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany. AD - Signaling Research Centers BIOSS and CIBSS, University of Freiburg, 79104 Freiburg, Germany. FAU - Prinz, Marco AU - Prinz M AD - Institute of Neuropathology, Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany. AD - Center for Basics in NeuroModulation (NeuroModulBasics), Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany. AD - Signaling Research Centers BIOSS and CIBSS, University of Freiburg, 79104 Freiburg, Germany. FAU - Kierdorf, Katrin AU - Kierdorf K AD - Institute of Neuropathology, Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany. AD - CIBSS-Center for Integrative Biological Signaling Studies, University of Freiburg, 79104 Freiburg, Germany. AD - Center for Basics in NeuroModulation (NeuroModulBasics), Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20220726 PL - United States TA - Cell Rep Methods JT - Cell reports methods JID - 9918227360606676 MH - Cell Differentiation/genetics MH - *Microphysiological Systems MH - Macrophages MH - Histiocytes MH - *Induced Pluripotent Stem Cells PMC - PMC9421540 OTO - NOTNLM OT - Kupffer cells OT - alveolar macrophages OT - bone marrow-derived macrophages OT - macrophages OT - microglia OT - organotypic cell culture OT - peritoneal macrophages OT - tissue normoxia OT - tissue-resident macrophages COIS- The authors declare no competing interests. EDAT- 2022/09/02 06:00 MHDA- 2022/09/02 06:01 PMCR- 2022/07/26 CRDT- 2022/09/01 02:32 PHST- 2021/07/19 00:00 [received] PHST- 2022/02/11 00:00 [revised] PHST- 2022/07/06 00:00 [accepted] PHST- 2022/09/01 02:32 [entrez] PHST- 2022/09/02 06:00 [pubmed] PHST- 2022/09/02 06:01 [medline] PHST- 2022/07/26 00:00 [pmc-release] AID - S2667-2375(22)00133-3 [pii] AID - 100260 [pii] AID - 10.1016/j.crmeth.2022.100260 [doi] PST - epublish SO - Cell Rep Methods. 2022 Jul 26;2(8):100260. doi: 10.1016/j.crmeth.2022.100260. eCollection 2022 Aug 22.