title: Highly angiogenic CXCR4(+)CD31(+) monocyte subset derived from 3D culture of human peripheral blood.
abstract: Ex vivo expansion of human circulating angiogenic cells is a major challenge in autologous cell therapy for ischemic diseases. Here, we demonstrate that hematosphere-derived CXCR4(+)CD31(+) myeloid cells using peripheral blood possess robust proangiogenic capacity such as formation of vessel-like structures and tip cell-like morphology in Matrigel. We also found that CD31 positive myeloid cells are principal cellular component of hematospheres by magnetic cell sorting. Flow cytometry analysis showed that fresh peripheral blood contained 40.3 ± 15.2% of CXCR4(+)CD31(+) myeloid cells, but at day 5 of hematosphere culture, most of myeloid cells were CXCR4(+)CD31(+) by 86.9 ± 5.4%. Hematosphere culture significantly increased the production of angiogenic niche-supporting cytokines. Moreover, CD31-homophilic interaction and VEGF-VEGF receptor loop signaling were essential for sphere formation and acquisition of angiogenic capacity in hematospheres. Matrigel plug and ischemic hindlimb model provide in vivo evidence that hematosphere-derived myeloid cells have highly vasculogenic capacities, participate in new and mature vessel formation, and exert therapeutic effects on ischemic hindlimb. In conclusion, our strategy for ex vivo expansion of human CXCR4(+)CD31(+) angiogenic cells using hematospheres provides an autologous therapeutic cell source for ischemic diseases and a new model for investigating the microenvironment of angiogenesis.
authors.affiliation: National Research Laboratory for Stem Cell Niche, Republic of Korea.
authors.forename: Jin
authors.initials: J
authors.lastname: Hur
authors.forename: Jae-Il
authors.initials: JI
authors.lastname: Choi
authors.forename: Ji-Yeon
authors.initials: JY
authors.lastname: Yun
authors.forename: Chang-Hwan
authors.initials: CH
authors.lastname: Yoon
authors.forename: Jae Hee
authors.initials: JH
authors.lastname: Jang
authors.forename: Seung-Gyun
authors.initials: SG
authors.lastname: Im
authors.forename: Seung-Bum
authors.initials: SB
authors.lastname: Ko
authors.forename: Jin-A
authors.initials: JA
authors.lastname: Kang
authors.forename: Jonghanne
authors.initials: J
authors.lastname: Park
authors.forename: Sang Eun
authors.initials: SE
authors.lastname: Lee
authors.forename: Ju-Young
authors.initials: JY
authors.lastname: Kim
authors.forename: Han-Mo
authors.initials: HM
authors.lastname: Yang
authors.forename: Young-Bae
authors.initials: YB
authors.lastname: Park
authors.forename: Hyo-Soo
authors.initials: HS
authors.lastname: Kim
mesh_terms: D000328:Adult; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D060833:Cellular Microenvironment / Q000187:drug effects; D003094:Collagen / Q000494:pharmacology; D004338:Drug Combinations; D006614:Hindlimb / Q000098:blood supply / Q000473:pathology; D006801:Humans; D007511:Ischemia / Q000473:pathology / Q000628:therapy; D007797:Laminin / Q000494:pharmacology; D018950:Lipopolysaccharide Receptors / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D009000:Monocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism* / Q000637:transplantation; D018919:Neovascularization, Physiologic / Q000187:drug effects; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D011509:Proteoglycans / Q000494:pharmacology; D019718:Receptors, CXCR4 / Q000378:metabolism*; D040262:Receptors, Vascular Endothelial Growth Factor / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D018874:Spheroids, Cellular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
chemical_list: D004338:Drug Combinations; D007797:Laminin; D018950:Lipopolysaccharide Receptors; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D011509:Proteoglycans; D019718:Receptors, CXCR4; D042461:Vascular Endothelial Growth Factor A; C059485:matrigel; D003094:Collagen; D040262:Receptors, Vascular Endothelial Growth Factor