Homing of purified murine lymphohematopoietic stem cells: a cytokine-induced defect.
This study was designed to establish a direct homing assay using purified lineage-negative Sca-1-positive (Lin(-) Sca(+)) murine bone marrow cells and to evaluate the effects of cytokines on homing. C57BL/6 Lin(-) Sca(+) marrow stem cells were labeled with 5-(and 6)-carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) and then injected by tail vein into untreated C57BL/6 mice. Marrow was harvested at various times after cell infusion and analyzed on a high-speed MoFlo cell sorter for fluorescent positive events, using a large event analysis, with at least 16 million total events analyzed. We have shown that homing of Lin(-) Sca(+) cells plateaus by 1 h, and at 3 h post-infusion is linear between 50,000 and 1,000,000 infused cells. This forms a base for a homing assay in which 250,000 CFDA-SE labeled Lin(-) Sca(+) marrow cells are infused and then recovered from marrow 3 h later, followed by a large-event fluorescence-activated cell sorting (FACS) analysis. We found that 7.45-9.32% of infused cells homed and that homing of stem cells cultured for 48 h in interleukin-3 (IL-3), IL-6, IL-11, and steel factor cultured cells was defective when compared to noncultured cells. Exposure of marrow stem cells to IL-3, IL-6, IL-11, and steel factor induces a stem cell homing defect, which probably underlies the engraftment defect previously characterized under these conditions.
Cerny
Jan
J
Department of Research, Roger Williams Medical Center, Providence, RI 02908, USA.
Dooner
Mark
M
McAuliffe
Christina
C
Habibian
Houri
H
Stencil
Kimberly
K
Berrios
Virla
V
Reilly
Judy
J
Carlson
Jane
J
Cerny
Anna Maria
AM
d'Hondt
Lionel
L
Benoit
Brian
B
Lambert
Jean-Francois
JF
Colvin
Gerald
G
Nilsson
Susan
S
Becker
Pamela
P
Quesenberry
Peter
P
D000818:Animals; D000943:Antigens, Differentiation / Q000032:analysis; D001854:Bone Marrow Cells / Q000166:cytology; D016026:Bone Marrow Transplantation / Q000379:methods*; D018929:Cell Culture Techniques / Q000379:methods; D002465:Cell Movement / Q000187:drug effects*; D016207:Cytokines / Q000494:pharmacology*; D006085:Graft Survival; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D017370:Interleukin-11 / Q000494:pharmacology; D007377:Interleukin-3 / Q000494:pharmacology; D015850:Interleukin-6 / Q000494:pharmacology; D007700:Kinetics; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D013154:Spleen / Q000166:cytology; D019089:Stem Cell Factor / Q000494:pharmacology; D013950:Thymus Gland / Q000166:cytology
D000943:Antigens, Differentiation; D016207:Cytokines; D017370:Interleukin-11; D007377:Interleukin-3; D015850:Interleukin-6; D019089:Stem Cell Factor; C047076:senescent cell differentiation antigen