Article
Leukemia Inhibitory Factor (LIF) Overexpression Increases the Angiogenic Potential of Bone Marrow Mesenchymal Stem/Stromal Cells
Registro en:
SANTOS, Girlaine Café et al. Leukemia Inhibitory Factor (LIF) Overexpression Increases the Angiogenic Potential of Bone Marrow Mesenchymal Stem/Stromal Cells. Frontiers in Cell and Developmental Biology, aug. 2020.
2296-634X
10.3389/fcell.2020.00778
Autor
Santos, Girlaine Café
Silva, Daniela Nascimento
Fortuna, Vitor
Silveira, Brysa Mariana
Orge, Iasmim Diniz
Santana, Thaís Alves de
Sampaio, Gabriela Louise
Paredes, Bruno Diaz
Santos, Ricardo Ribeiro dos
Soares, Milena Botelho Pereira
Resumen
Brazilian Research Council (CNPq)/MS grant
number 443909/2018-0. 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.