Application of polychromatic flow cytometry to identify novel subsets of circulating cells with angiogenic potential.
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.
Estes
Myka L
ML
Department of Pediatrics, Indiana University, Indianapolis, Indiana, USA.
Mund
Julie A
JA
Mead
Laura E
LE
Prater
Daniel N
DN
Cai
Shanbao
S
Wang
Haiyan
H
Pollok
Karen E
KE
Murphy
Michael P
MP
An
Caroline S T
CS
Srour
Edward F
EF
Ingram
David A
DA
Case
Jamie
J
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
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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