Characterization of CD34+ human hemopoietic progenitor cells from the peripheral blood: enzyme-, carbohydrate- and immunocytochemistry, morphometry, and ultrastructure.
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.
Thiele
J
J
Institute of Pathology, University of Cologne, Germany.
Wickenhauser
C
C
Baldus
S E
SE
Kuemmel
T
T
Zirbes
T K
TK
Drebber
U
U
Wirtz
R
R
Thiel
A
A
Hansmann
M L
ML
Fischer
R
R
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
D015703:Antigens, CD; D018952:Antigens, CD34; D002241:Carbohydrates; D004798:Enzymes; D008562:Membrane Glycoproteins; D009077:Mucins; D011975:Receptors, Mitogen; D052116:Sialomucins