Canine stem cell factor augments expression of matrix metalloproteinase-9 by CD34 cells.
BACKGROUND
Canine models have proved to be predictive of clinical findings in human bone marrow (BM) transplantation; consequently, the utilization of dogs is an excellent tool for supporting therapeutic purposes. Considering the role of growth factors in homing and mobilization of hematopoietic progenitors, the aim of this work was to evaluate whether canine stem cell factor (cSCF) contributes to matrix metalloproteinase (MMP)-9 secretion by CD34 cells.


METHODS
The study was carried out in a cell population selected by immunomagnetic techniques using the anti-canine CD34 monoclonal antibody (MAb) 3B4 produced by us. Secretion of MMP-9 was evaluated by zymography.


RESULTS
Analyzes of canine CD34(+) cells guaranteed that the MAb 3B4 was optimum for selecting a subset population with defined characteristics of primitive hematopoietic cells. The isolated cells were able to proliferate onto irradiated pre-established stroma, giving rise to mature neutrophils. There was also a 20-fold enrichment in the long-term culture-initiating cell content when the isolated population was added to irradiated cultures, with respect to the starting mononuclear cell population.


CONCLUSIONS
We have provided the first evidence that canine BM CD34(+) cells constitutively express MMP-9 and the role of cSCF in up-regulating the secretion of this enzyme. The fact that cSCF augments expression of MMP-9 together with the ability of the isolated CD34(+)cells to proliferate onto irradiated pre-established stroma enables further investigations to determine whether the secretion of MMP-9 mediated by cSCF is one of the factors that enhance migration, homing and repopulation of primitive hemopoietic cells.
Ostronoff
L K
LK
Departamento de Bioquímica y Biología Molecular, Facultad de Veterinaria, Universidad Complutense de Madrid, Madrid, Spain.
Kremmer
E
E
Fermín
M L
ML
Fragío
C
C
Mysliwietz
J
J
Kolb
H-J
HJ
Tejero
C
C
D000818:Animals; D000911:Antibodies, Monoclonal; D018952:Antigens, CD34 / Q000378:metabolism*; D015153:Blotting, Western; D002460:Cell Line; D049109:Cell Proliferation; D003114:Colony-Forming Units Assay; D004285:Dogs; D005260:Female; D005434:Flow Cytometry; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism*; D019089:Stem Cell Factor / Q000378:metabolism*; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D018952:Antigens, CD34; D019089:Stem Cell Factor; D020780:Matrix Metalloproteinase 9