title: In vitro modeling of endothelial interaction with macrophages and pericytes demonstrates Notch signaling function in the vascular microenvironment.
abstract: Angiogenesis is regulated by complex interactions between endothelial cells and support cells of the vascular microenvironment, such as tissue myeloid cells and vascular mural cells. Multicellular interactions during angiogenesis are difficult to study in animals and challenging in a reductive setting. We incorporated stromal cells into an established bead-based capillary sprouting assay to develop assays that faithfully reproduce major steps of vessel sprouting and maturation. We observed that macrophages enhance angiogenesis, increasing the number and length of endothelial sprouts, a property we have dubbed "angiotrophism." We found that polarizing macrophages toward a pro-inflammatory profile further increased their angiotrophic stimulation of vessel sprouting, and this increase was dependent on macrophage Notch signaling. To study endothelial/pericyte interactions, we added vascular pericytes directly to the bead-bound endothelial monolayer. These pericytes formed close associations with the endothelial sprouts, causing increased sprout number and vessel caliber. We found that Jagged1 expression and Notch signaling are essential for the growth of both endothelial cells and pericytes and may function in their interaction. We observed that combining endothelial cells with both macrophages and pericytes in the same sprouting assay has multiplicative effects on sprouting. These results significantly improve bead-capillary sprouting assays and provide an enhanced method for modeling interactions between the endothelium and the vascular microenvironment. Achieving this in a reductive in vitro setting represents a significant step toward a better understanding of the cellular elements that contribute to the formation of mature vasculature.
authors.affiliation: Obstetrics/Gynecology, Columbia University, New York, NY, USA.
authors.forename: Ian W
authors.initials: IW
authors.lastname: Tattersall
authors.affiliation: Obstetrics/Gynecology, Columbia University, New York, NY, USA.|Pathology and Cell Biology, Columbia University, New York, NY, USA.
authors.forename: Jing
authors.initials: J
authors.lastname: Du
authors.affiliation: Obstetrics/Gynecology, Columbia University, New York, NY, USA.
authors.forename: Zhuangzhuang
authors.initials: Z
authors.lastname: Cong
authors.affiliation: Obstetrics/Gynecology, Columbia University, New York, NY, USA.
authors.forename: Bennet S
authors.initials: BS
authors.lastname: Cho
authors.affiliation: Obstetrics/Gynecology, Columbia University, New York, NY, USA.
authors.forename: Alyssa M
authors.initials: AM
authors.lastname: Klein
authors.affiliation: Obstetrics/Gynecology, Columbia University, New York, NY, USA.
authors.forename: Chelsea L
authors.initials: CL
authors.lastname: Dieck
authors.affiliation: Obstetrics/Gynecology, Columbia University, New York, NY, USA.
authors.forename: Reyhaan A
authors.initials: RA
authors.lastname: Chaudhri
authors.affiliation: Obstetrics/Gynecology, Columbia University, New York, NY, USA.
authors.forename: Henar
authors.initials: H
authors.lastname: Cuervo
authors.affiliation: Obstetrics/Gynecology, Columbia University, New York, NY, USA.
authors.forename: James H
authors.initials: JH
authors.lastname: Herts
authors.affiliation: Obstetrics/Gynecology, Columbia University, New York, NY, USA. kitaj@uic.edu.|Pathology and Cell Biology, Columbia University, New York, NY, USA. kitaj@uic.edu.|Physiology and Biophysics, University of Illinois Chicago, College of Medicine, Chicago, IL, USA. kitaj@uic.edu.|Columbia University Medical Center, 1130 St. Nicholas Avenue, ICRC 926, New York, NY, 10032, USA. kitaj@uic.edu.
authors.forename: Jan
authors.initials: J
authors.lastname: Kitajewski
mesh_terms: D000818:Animals; D002450:Cell Communication; D002460:Cell Line; D016764:Cell Polarity; D002470:Cell Survival; D060833:Cellular Microenvironment; D018920:Coculture Techniques; D055785:Gene Knockdown Techniques; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D007249:Inflammation / Q000473:pathology; D018836:Inflammation Mediators / Q000378:metabolism; D008264:Macrophages / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D008954:Models, Biological; D022423:Myeloid Cells / Q000166:cytology / Q000378:metabolism; D018919:Neovascularization, Physiologic; D020286:Pericytes / Q000166:cytology* / Q000378:metabolism; D051880:Receptors, Notch / Q000378:metabolism*; D000072099:Serrate-Jagged Proteins / Q000378:metabolism; D015398:Signal Transduction
chemical_list: D018836:Inflammation Mediators; D051880:Receptors, Notch; D000072099:Serrate-Jagged Proteins
keywords: Endothelial cell; In vitro; Macrophage; Notch; Pericyte