title: Putative CD133+ melanoma cancer stem cells induce initial angiogenesis in vivo.
abstract: Tumor angiogenesis is essential for tumor growth and metastasis, and is regulated by a complex network of various types of cells, chemokines, and stimulating factors. In contrast to sprouting angiogenesis, tumor angiogenesis is also influenced by hypoxia, inflammation, and the attraction of bone-marrow-derived cells. Recently, cancer stem cells have been reported to mimic vascularization by differentiating into endothelial cells and inducing vessel formation. In this study, the influence of cancer stem cells on initial angiogenesis was evaluated for the metastatic melanoma cell line D10. Following flow cytometry, CD133+ and CD133- cells were isolated using magnetic cell separation and different cell fractions were transferred to porcine gelatin sponges, which were implanted into the dorsal skinfold chamber of immunocompromised mice. Angiogenesis was analyzed based on microvessel density over a 10-day period using in vivo fluorescence microscopy, and the results were verified using immunohistology. CD133+ D10 cells showed a significant induction of early angiogenesis in vivo, contrary to CD133- D10 cells, unsorted D10 cells, and negative control. Neovascularization was confirmed by visualizing endothelial cells by immunohistology using an anti-CD31 antibody. Because CD133+ cells are rare in clinical specimens and hardly amenable to functional assays, the D10 cell line provides a suitable model to study the angiogenic potential of putative cancer stem cells and the leukocyte-endothelial cell interaction in the dorsal skinfold chamber in vivo. This cancer stem cell model might be useful in the development and evaluation of therapeutic agents targeting tumors.
authors.affiliation: Department of Oral and Maxillofacial Surgery, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany. Electronic address: zimmerer.ruediger@mh-hannover.de.
authors.forename: Rüdiger M
authors.initials: RM
authors.lastname: Zimmerer
authors.affiliation: Department of Oral and Maxillofacial Surgery, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany. Electronic address: pjmatthiesen@web.de.
authors.forename: Peter
authors.initials: P
authors.lastname: Matthiesen
authors.affiliation: Department of Oral and Maxillofacial Surgery, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany. Electronic address: kreher.fritjof@mh-hannover.de.
authors.forename: Fritjof
authors.initials: F
authors.lastname: Kreher
authors.affiliation: Department of Oral and Maxillofacial Surgery, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany. Electronic address: kampmann.andreas@mh-hannover.de.
authors.forename: Andreas
authors.initials: A
authors.lastname: Kampmann
authors.affiliation: Department of Oral and Maxillofacial Surgery, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany. Electronic address: spalthoff.simon@mh-hannonver.de.
authors.forename: Simon
authors.initials: S
authors.lastname: Spalthoff
authors.affiliation: Department of Oral and Maxillofacial Surgery, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany. Electronic address: jehn.philipp@mh-hannover.de.
authors.forename: Philipp
authors.initials: P
authors.lastname: Jehn
authors.affiliation: Department of Oral and Maxillofacial Surgery, University of Freiburg Medical School, Hugstetter Str. 53, 70164 Freiburg, Germany. Electronic address: bittermann.guido@uniklinik-freiburg.de.
authors.forename: Gido
authors.initials: G
authors.lastname: Bittermann
authors.affiliation: Department of Oral and Maxillofacial Surgery, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany. Electronic address: gellrich.nils-claudius@mh-hannover.de.
authors.forename: Nils-Claudius
authors.initials: NC
authors.lastname: Gellrich
authors.affiliation: Department of Oral and Maxillofacial Surgery, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany. Electronic address: tavassol.frank@mh-hannover.de.
authors.forename: Frank
authors.initials: F
authors.lastname: Tavassol
mesh_terms: D000071916:AC133 Antigen; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism*; D045744:Cell Line, Tumor; D005260:Female; D006023:Glycoproteins / Q000378:metabolism*; D006439:Hemodynamics; D006801:Humans; D018189:Immunomagnetic Separation; D000069416:Intravital Microscopy; D008545:Melanoma / Q000098:blood supply* / Q000276:immunology / Q000473:pathology*; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D008856:Microscopy, Fluorescence; D055806:Microvessels / Q000473:pathology / Q000503:physiopathology; D014411:Neoplastic Stem Cells / Q000276:immunology* / Q000473:pathology*; D009389:Neovascularization, Pathologic; D010455:Peptides / Q000378:metabolism*
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse
keywords: Angiogenesis; CD133; Cancer stem cells; Dorsal skinfold chamber; In vivo fluorescence microscopy; Melanoma