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https://www.arca.fiocruz.br/handle/icict/45741
LEUKEMIA INHIBITORY FACTOR (LIF) OVEREXPRESSION INCREASES THE ANGIOGENIC POTENTIAL OF BONE MARROW MESENCHYMAL STEM/STROMAL CELLS
Genética
Receptor de Fator Inibidor de Leucemia
Angiogênese
Genetic modification
LIF
Proangiogenico factors
Angiogenesis
Author
Affilliation
Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Bahia, Brasil / Health Institute of Technology. Salvador, BA, Brazil.
Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Bahia, Brasil / Health Institute of Technology. Salvador, BA, Brazil.
Federal University of Bahia. Health Sciences Institute. Salvador, BA, Brazil.
Federal University of Bahia. Health Sciences Institute. Salvador, BA, Brazil.
Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Bahia, Brasil / Health Institute of Technology. Salvador, BA, Brazil.
Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Bahia, Brasil / Health Institute of Technology. Salvador, BA, Brazil.
Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Bahia, Brasil.
Research D’Or Institute. Rio de Janeiro, RJ, Brazil.
Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Bahia, Brasil / Health Institute of Technology. Salvador, BA, Brazil / ational Institute of Science and Technology for Regenerative Medicine. Rio de Janeiro, RJ, Brazil.
Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Bahia, Brasil / Health Institute of Technology. Salvador, BA, Brazil / ational Institute of Science and Technology for Regenerative Medicine. Rio de Janeiro, RJ, Brazil.
Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Bahia, Brasil / Health Institute of Technology. Salvador, BA, Brazil.
Federal University of Bahia. Health Sciences Institute. Salvador, BA, Brazil.
Federal University of Bahia. Health Sciences Institute. Salvador, BA, Brazil.
Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Bahia, Brasil / Health Institute of Technology. Salvador, BA, Brazil.
Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Bahia, Brasil / Health Institute of Technology. Salvador, BA, Brazil.
Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Bahia, Brasil.
Research D’Or Institute. Rio de Janeiro, RJ, Brazil.
Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Bahia, Brasil / Health Institute of Technology. Salvador, BA, Brazil / ational Institute of Science and Technology for Regenerative Medicine. Rio de Janeiro, RJ, Brazil.
Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Bahia, Brasil / Health Institute of Technology. Salvador, BA, Brazil / ational Institute of Science and Technology for Regenerative Medicine. Rio de Janeiro, RJ, Brazil.
Abstract
Mesenchymal stem/stromal cells (MSCs) have the ability to secrete bioactive
molecules, exerting multiple biological effects, such as tissue regeneration, reduction
of inflammation, and neovascularization. The therapeutic potential of MSCs can be
increased by genetic modification to overexpress cytokines and growth factors. Here
we produced mouse MSCs overexpressing human leukemia inhibitory factor (LIF) to
assess their proangiogenic potential in vitro and in vivo. Mouse bone marrow-derived
MSCs were transduced by using a second-generation lentiviral system to express
human LIF. Leukemia inhibitory factor expression was confirmed by RT-qPCR and by
ELISA, allowing the quantification of the transcript and secreted protein, respectively.
Flow cytometry analysis and trilineage differentiation assay showed that the MSC_LIF
cell line maintained the immunophenotype and a multipotency characteristic of MSCs.
The immunosuppressive activity of MSC_LIF was confirmed using a lymphoproliferation
assay. Moreover, gene expression analysis demonstrated upregulation of genes coding
for strategic factors in the neovascularization process, such as angiogenin, IL-8,
MCP-1, and VEGF, and for the perivascular cell markers aSMA, Col4a1, SM22,
and NG2. To evaluate the pro-angiogenic potential of MSC_LIF, we first tested its
effects on endothelial cells obtained from umbilical vein in a scratch wound healing
assay. Conditioned medium (CM) from MSC_LIF promoted a significant increase in cell
migration compared to CM from control MSC. Additionally, in vitro tube formation of
endothelial cells was increased by the presence of MSC_LIF, as shown in microvessel
sprouting in aortic ring cultures. Finally, an in vivo Matrigel plug assay was performed,
showing that MSC_LIF were more potent in promoting in vivo angiogenesis and tissue
vascularization than control MSCs. In conclusion, LIF overexpression is a promising
strategy to increase the proangiogenic potential of MSCs and sets precedents for future
investigations of their potential applications for the treatment of ischemic diseases and
tissue repair.
Keywords in Portuguese
Células-tronco mesenquimaisGenética
Receptor de Fator Inibidor de Leucemia
Angiogênese
Keywords
Mesenchymal stem/stromal cellsGenetic modification
LIF
Proangiogenico factors
Angiogenesis
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