Homing and clonogenic outgrowth of CD34(+) peripheral blood stem cells: a role for L-selectin?
OBJECTIVE
After transplantation of hematopoietic stem cells, adhesion molecules play a major role in the multistep process of engraftment in which L-selectin is suggested to be of relevance. A positive correlation previously was found between the number of reinfused L-selectin(+) stem cells and platelet recovery. In the present study, we determined the role of L-selectin in different engraftment steps, i.e., adhesion to endothelial cells, migration, and clonogenic outgrowth by in vitro assays that closely mimic the in vivo situation.


METHODS
Flow adhesion and migration experiments were performed using the human bone marrow endothelial cell line 4LHBMEC and isolated peripheral CD34(+) cells with or without blocking of L-selectin-ligand interaction. Various clonogenic assays, including serum-free colony-forming unit-megakaryocytes (CFU-MK) and burst-forming unit-megakaryocytes (BFU-MK), were performed with sorted L-selectin(+)L-selectin(-) cells or in the presence of antibodies.


RESULTS
Blocking of L-selectin on CD34(+) cells did not significantly affect rolling over and firm adhesion to 4LHBMEC. In addition, no role for L-selectin was found in transendothelial migration experiments. Finally, in clonogenic outgrowth of sorted or anti-L-selectin monoclonal antibody-incubated CD34(+) cells, no key role for L-selectin expression could be defined in BFU-MK and CFU-MK assays.


CONCLUSIONS
Using in vitro assays for CD34(+) stem cell adhesion, migration, and clonogenic capacity, we were not able to define a major role for L-selectin.
de Boer
Fransien
F
Department of Hematology, VU University Medical Center, Amsterdam, The Netherlands.
Kessler
Floortje L
FL
Netelenbos
Tanja
T
Zweegman
Sonja
S
Huijgens
Peter C
PC
van der Wall
Elsken
E
van der Linden
Jan A M
JA
Pinedo
Herbert M
HM
Schuurhuis
Gerrit Jan
GJ
Dräger
Angelika M
AM
D015703:Antigens, CD / Q000097:blood; D018952:Antigens, CD34 / Q000097:blood*; D001854:Bone Marrow Cells / Q000166:cytology; D001943:Breast Neoplasms / Q000097:blood / Q000276:immunology; D002448:Cell Adhesion; D002454:Cell Differentiation; D002455:Cell Division; D002460:Cell Line; D002465:Cell Movement; D003114:Colony-Forming Units Assay; D004730:Endothelium, Vascular / Q000502:physiology; D005260:Female; D006412:Hematopoietic Stem Cells / Q000473:pathology*; D006801:Humans; D019041:L-Selectin / Q000502:physiology*; D008223:Lymphoma / Q000097:blood / Q000276:immunology; D008533:Megakaryocytes / Q000473:pathology; D009101:Multiple Myeloma / Q000097:blood / Q000276:immunology; D014410:Tumor Stem Cell Assay
D015703:Antigens, CD; D018952:Antigens, CD34; D019041:L-Selectin