title: Application of polychromatic flow cytometry to identify novel subsets of circulating cells with angiogenic potential.
abstract: Defining whether human circulating proangiogenic cells represent a subset of the hematopoietic system and express CD45 or are hematopoietic derivatives that do not express CD45 (and are called endothelial progenitor cells) remains controversial. We have previously developed a polychromatic flow cytometry (PFC) protocol to isolate subsets of hematopoietic cells and we now identify the circulating pool of CD34(+)CD45(dim) cells representing functional circulating hematopoietic stem and progenitor cells (CHSPCs) that can be separated on the basis of AC133 expression and report that the AC133(+) subset of the CHSPCs enhances the growth of tumor blood vessels in vivo in immunodeficient mice. In addition, the ratio of AC133(+) proangiogenic CHSPCs to AC133(-) nonangiogenic CHSPCs unambiguously correlates with the severity of the clinical state of patients with peripheral arterial disease. In sum, a PFC protocol validated via in vitro and in vivo analyses, can be used to interrogate the roles of human hematopoietic elements in the growth and maintenance of the vasculature.
authors.affiliation: Department of Pediatrics, Indiana University, Indianapolis, Indiana, USA.
authors.forename: Myka L
authors.initials: ML
authors.lastname: Estes
authors.forename: Julie A
authors.initials: JA
authors.lastname: Mund
authors.forename: Laura E
authors.initials: LE
authors.lastname: Mead
authors.forename: Daniel N
authors.initials: DN
authors.lastname: Prater
authors.forename: Shanbao
authors.initials: S
authors.lastname: Cai
authors.forename: Haiyan
authors.initials: H
authors.lastname: Wang
authors.forename: Karen E
authors.initials: KE
authors.lastname: Pollok
authors.forename: Michael P
authors.initials: MP
authors.lastname: Murphy
authors.forename: Caroline S T
authors.initials: CS
authors.lastname: An
authors.forename: Edward F
authors.initials: EF
authors.lastname: Srour
authors.forename: David A
authors.initials: DA
authors.lastname: Ingram
authors.forename: Jamie
authors.initials: J
authors.lastname: Case
mesh_terms: D000071916:AC133 Antigen; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D015703:Antigens, CD / Q000032:analysis; D018952:Antigens, CD34 / Q000032:analysis; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D006023:Glycoproteins / Q000032:analysis; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D007231:Infant, Newborn; D017493:Leukocyte Common Antigens / Q000032:analysis; D008297:Male; D051379:Mice; D016513:Mice, SCID; D008875:Middle Aged; D009000:Monocytes / Q000166:cytology; D009389:Neovascularization, Pathologic / Q000473:pathology; D010455:Peptides / Q000032:analysis; D058729:Peripheral Arterial Disease / Q000473:pathology*; D012720:Severity of Illness Index; D013194:Staining and Labeling / Q000379:methods*; D055815:Young Adult
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D018952:Antigens, CD34; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse; D017493:Leukocyte Common Antigens
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