title: Characterization of CD34+ human hemopoietic progenitor cells from the peripheral blood: enzyme-, carbohydrate- and immunocytochemistry, morphometry, and ultrastructure.
abstract: Following an immunomagnetic isolation and enrichment procedure, CD34+ cells were harvested from the peripheral blood of about 50 healthy donors. A battery of cytochemical staining reactions, monoclonal and carbohydrate-specific antibodies, proliferation markers and lectins was applied on smears and sections from paraffin-embedded pellets. Additionally, a morphometric analysis and ultrastructural investigation was carried out. More than 95% of the total yield of progenitor cells expressed CD34 and CD43 (MT1) and of these about 90% CD45 (LCA) and 25% CD45A (MT2). The CD34+/CD45RA-population was thought to represent very primitive, probably not lineage-restricted stem cells. On the other hand, reactivity with ANAE, CD11c, CD15, CD20, Ret40f, KiM1P, and CD61 (ranging between 1 to 20%) was considered to indicate a transition into more differentiated elements of hemopoiesis. The failure to detect any staining with proliferation markers (Ki-67/MIB 1, PCNA, KiS1) was in keeping with a quiescent status. Carbohydrate antigens revealed a pattern which underlines the fact that the CD34 and CD43 antigens belong to the family of heavily O-glycosylated sialomucins. Blood group antigens which are located at the peripheral regions of mucin-oligosaccharides (H type 2, Lewis, Lewis) could be demonstrated, but not A, B, Sialyl-Lewis and Lewis. Morphometric analysis revealed that CD34+ progenitors were larger than small lymphocytes. Electron microscopy showed a relatively primitive cytoplasmic organization and numerous tiny magnetic beads clustered at the plasma membrane.
authors.affiliation: Institute of Pathology, University of Cologne, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Thiele
authors.forename: C
authors.initials: C
authors.lastname: Wickenhauser
authors.forename: S E
authors.initials: SE
authors.lastname: Baldus
authors.forename: T
authors.initials: T
authors.lastname: Kuemmel
authors.forename: T K
authors.initials: TK
authors.lastname: Zirbes
authors.forename: U
authors.initials: U
authors.lastname: Drebber
authors.forename: R
authors.initials: R
authors.lastname: Wirtz
authors.forename: A
authors.initials: A
authors.lastname: Thiel
authors.forename: M L
authors.initials: ML
authors.lastname: Hansmann
authors.forename: R
authors.initials: R
authors.lastname: Fischer
mesh_terms: D000328:Adult; D015703:Antigens, CD / Q000032:analysis; D018952:Antigens, CD34; D001773:Blood Cells; D002241:Carbohydrates / Q000032:analysis; D002455:Cell Division; D004798:Enzymes / Q000032:analysis; D006412:Hematopoietic Stem Cells / Q000276:immunology / Q000378:metabolism / Q000648:ultrastructure; D006801:Humans; D018189:Immunomagnetic Separation; D008562:Membrane Glycoproteins / Q000032:analysis; D009077:Mucins / Q000032:analysis; D011975:Receptors, Mitogen / Q000032:analysis; D052116:Sialomucins
chemical_list: D015703:Antigens, CD; D018952:Antigens, CD34; D002241:Carbohydrates; D004798:Enzymes; D008562:Membrane Glycoproteins; D009077:Mucins; D011975:Receptors, Mitogen; D052116:Sialomucins