Huaning Zhao1,2, Jordan Darden1,3, John C Chappell1,2,3,4. 1. Center for Heart and Regenerative Medicine, Virginia Tech Carilion Research Institute, Roanoke, VA, USA. 2. Department of Biomedical Engineering and Mechanics, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA. 3. Graduate Program in Translational Biology, Medicine, and Health, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA. 4. Department of Basic Science Education, Virginia Tech Carilion School of Medicine, Roanoke, VA, USA.
Abstract
OBJECTIVE: Pericytes are specialized perivascular cells embedded within the basement membrane. These cells envelope the abluminal surface of endothelial cells and promote microvessel homeostasis. Recent discoveries of unique pericyte functions, particularly in neural tissues, underscore the need for overcoming existing challenges in establishing a functionally validated pericyte cell line. Here, we present methodologies for addressing these challenges as well as an embryonic pericyte cell line for use with in vitro and ex vivo experimental models. METHODS: We isolated an enriched population of NG2:DsRed+ pericytes from E12.5 mice. This pericyte cell line was compared to MEFs with respect to gene expression, cell morphology and migration, and engagement with endothelial cells during junction stabilization and angiogenesis. RESULTS: NG2+ pericytes displayed gene expression patterns, cell morphology, and 2D migration behaviors distinct from MEFs. In three different vessel formation models, pericytes from this line migrated to and incorporated into developing vessels. When co-cultured with HUVECs, these pericytes stimulated more robust VE-Cadherin junctions between HUVECs as compared to MEFs, as well as contributed to HUVEC organization into primitive vascular structures. CONCLUSIONS: Our data support use of this pericyte cell line in a broad range of models to further understand pericyte functionality during normal and pathological conditions.
OBJECTIVE: Pericytes are specialized perivascular cells embedded within the basement membrane. These cells envelope the abluminal surface of endothelial cells and promote microvessel homeostasis. Recent discoveries of unique pericyte functions, particularly in neural tissues, underscore the need for overcoming existing challenges in establishing a functionally validated pericyte cell line. Here, we present methodologies for addressing these challenges as well as an embryonic pericyte cell line for use with in vitro and ex vivo experimental models. METHODS: We isolated an enriched population of NG2:DsRed+ pericytes from E12.5 mice. This pericyte cell line was compared to MEFs with respect to gene expression, cell morphology and migration, and engagement with endothelial cells during junction stabilization and angiogenesis. RESULTS:NG2+ pericytes displayed gene expression patterns, cell morphology, and 2D migration behaviors distinct from MEFs. In three different vessel formation models, pericytes from this line migrated to and incorporated into developing vessels. When co-cultured with HUVECs, these pericytes stimulated more robust VE-Cadherin junctions between HUVECs as compared to MEFs, as well as contributed to HUVEC organization into primitive vascular structures. CONCLUSIONS: Our data support use of this pericyte cell line in a broad range of models to further understand pericyte functionality during normal and pathological conditions.
Authors: Jennifer T Durham; Brian M Dulmovits; Stephen M Cronk; Anthony R Sheets; Ira M Herman Journal: Invest Ophthalmol Vis Sci Date: 2015-06 Impact factor: 4.799
Authors: Johannes Schindelin; Ignacio Arganda-Carreras; Erwin Frise; Verena Kaynig; Mark Longair; Tobias Pietzsch; Stephan Preibisch; Curtis Rueden; Stephan Saalfeld; Benjamin Schmid; Jean-Yves Tinevez; Daniel James White; Volker Hartenstein; Kevin Eliceiri; Pavel Tomancak; Albert Cardona Journal: Nat Methods Date: 2012-06-28 Impact factor: 28.547
Authors: John C Chappell; Julia G Cluceru; Jessica E Nesmith; Kevin P Mouillesseaux; Vanessa B Bradley; Caitlin M Hartland; Yasmin L Hashambhoy-Ramsay; Joseph Walpole; Shayn M Peirce; Feilim Mac Gabhann; Victoria L Bautch Journal: Cardiovasc Res Date: 2016-05-03 Impact factor: 10.787
Authors: Laura B Payne; Huaning Zhao; Carissa C James; Jordan Darden; David McGuire; Sarah Taylor; James W Smyth; John C Chappell Journal: Microcirculation Date: 2019-05-27 Impact factor: 2.628
Authors: Natalia Kosyakova; Derek D Kao; Maria Figetakis; Francesc López-Giráldez; Susann Spindler; Morven Graham; Kevin J James; Jee Won Shin; Xinran Liu; Gregory T Tietjen; Jordan S Pober; William G Chang Journal: NPJ Regen Med Date: 2020-01-06
Authors: Natalia Kosyakova; Derek D Kao; Maria Figetakis; Francesc López-Giráldez; Susann Spindler; Morven Graham; Kevin J James; Jee Won Shin; Xinran Liu; Gregory T Tietjen; Jordan S Pober; William G Chang Journal: NPJ Regen Med Date: 2020-01-06