IFIBYNE   05513
INSTITUTO DE FISIOLOGIA, BIOLOGIA MOLECULAR Y NEUROCIENCIAS
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Gene expression of hematopoietic stimulating cytokines in human Adipose Derived Mesenchymal Stem Cells (hASC) sensitive and insensitive to TGF-β
Autor/es:
RODRIGUEZ, TM; CARREA, A; AVEDILLO DIEZ, I; SALDIAS, A; GALLO, R; PERONE, MJ; DEWEY, RA
Lugar:
Buenos Aires
Reunión:
Simposio; International Symposium on Stem Cell Research; 2009
Resumen:
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Human adipose stem cells
(hASC) have arisen as a safe and abundant source of stem cells for regenerative
medicine. Although they share many characteristics with bone marrow derived
mesenchymal stem cells (hBM-MSC), they remain much less characterized. It is known that hBM-MSCs secrete hematopoietic stem
and progenitor cells (HSPC) stimulating factors, but in hASCs these factors
have not been characterized in detail yet. Transforming Growth Factor (TGF)-b modulates multiple biological functions including the expression of
HSPC-stimulating cytokines by stromal cells. TGF-b1 maintains primitive
hematopoietic stem/progenitor cells in a quiescent or slow cycling state, in
part by down-regulating the expression of mitogenic cytokines. TGF-β1 responses are basically mediated by Type I (TβRI) and II (TβRII) surface receptors, which are expressed in
most cell types. Here we show the biological effect of inhibition of the TGF-β cascade on the expression pattern of hematopoietic stimulating cytokines
in hASC. To achieve this, we designed lentiviral vectors encoding two different
TGF-b receptor II (TβRII) dominant negative mutants (DN): TbRIIA-DN and TbRIIB-DN; and with them, we transduced purified and characterized hASC from
lipoaspirates. Activity of lentiviral encoded DN mutants was determined by MTT
assay on the TGF-b sensitive Mv1-Lu mink cell line. Expression
levels of Stem Cell Factor (SCF) (membrane bound and soluble), IL-6,
Thrombopoietin (TPO) and Flt3-ligand (FL) were studied by semi-quantitative
RT-PCR in TGF-β sensitive and insensitive cells. We found that hASC express either soluble
or membrane bound SCF, but not TPO. Additionally, we observed that, TGF-β negatively modulates SCF and FL expression, and positively IL-6. We also
detected that in genetically-modified hASC by lentiviral vectors expressing
only the marker transgene GFP, the expression levels of all studied cytokines
are significantly altered. A potential clinical application of hASC is to
exploit their ability to secrete early acting cytokines (FL, SCF and TPO) to
expand hemopoietic progenitor cells ex
vivo. Our results suggest that to obtain full HSPC expansion ex vivo from hASC, TPO expression should
be up-regulated and TGF-b action must be avoided.