Therapeutic angiogenesis induced by human umbilical cord tissue-derived mesenchymal stromal cells in a murine model of hindlimb ischemia.
BACKGROUND
Mesenchymal stem cells derived from human umbilical cord tissue, termed UCX®, have the potential to promote a full range of events leading to tissue regeneration and homeostasis. The main goal of this work was to investigate UCX® action in experimentally induced hindlimb ischemia (HLI).


METHODS
UCX®, obtained by using a proprietary technology developed by ECBio (Amadora, Portugal), were delivered via intramuscular injection to C57BL/6 females after unilateral HLI induction. Perfusion recovery, capillary and collateral density increase were evaluated by laser doppler, CD31 immunohistochemistry and diaphonisation, respectively. The activation state of endothelial cells (ECs) was analysed after EC isolation by laser capture microdissection microscopy followed by RNA extraction, cDNA synthesis and quantitative RT-PCR analysis. The UCX®-conditioned medium was analysed on Gallios flow cytometer. The capacity of UCX® in promoting tubulogenesis and EC migration was assessed by matrigel tubule formation and wound-healing assay, respectively.


RESULTS
We demonstrated that UCX® enhance angiogenesis in vitro via a paracrine effect. Importantly, after HLI induction, UCX® improve blood perfusion by stimulating angiogenesis and arteriogenesis. This is achieved through a new mechanism in which durable and simultaneous upregulation of transforming growth factor β2, angiopoietin 2, fibroblast growth factor 2, and hepatocyte growth factor, in endothelial cells is induced by UCX®.


CONCLUSIONS
In conclusion, our data demonstrate that UCX® improve the angiogenic potency of endothelial cells in the murine ischemic limb suggesting the potential of UCX® as a new therapeutic tool for critical limb ischemia.
Centro Cardiovascular da Universidade de Lisboa, Av. Prof Egas Moniz, 1649-028, Lisbon, Portugal.
Ana Rita S
AR
Pereira
Centro Cardiovascular da Universidade de Lisboa, Av. Prof Egas Moniz, 1649-028, Lisbon, Portugal.
Teresa F
TF
Mendes
Centro Cardiovascular da Universidade de Lisboa, Av. Prof Egas Moniz, 1649-028, Lisbon, Portugal.|Centro Hospitalar Lisboa Norte, Av. Prof. Egas Moniz, 1649-035, Lisbon, Portugal.
Augusto
A
Ministro
ECBio, Investigação e Desenvolvimento em Biotecnologia S.A., R. Henrique Paiva Couceiro, 27, 2700-4511, Amadora, Portugal.
Mariana
M
Teixeira
ECBio, Investigação e Desenvolvimento em Biotecnologia S.A., R. Henrique Paiva Couceiro, 27, 2700-4511, Amadora, Portugal.
Mariana
M
Filipe
ECBio, Investigação e Desenvolvimento em Biotecnologia S.A., R. Henrique Paiva Couceiro, 27, 2700-4511, Amadora, Portugal.
Jorge M
JM
Santos
ECBio, Investigação e Desenvolvimento em Biotecnologia S.A., R. Henrique Paiva Couceiro, 27, 2700-4511, Amadora, Portugal.
Rita N
RN
Bárcia
Nova Medical School/Faculdade de Ciências Médicas, Universidade Nova de Lisboa, 1169-056, Lisbon, Portugal.
J
J
Goyri-O'Neill
Centro Cardiovascular da Universidade de Lisboa, Av. Prof Egas Moniz, 1649-028, Lisbon, Portugal.|Faculdade de Medicina da Universidade de Lisboa, Av. Prof Egas Moniz, 1649-028, Lisbon, Portugal.
Fausto
F
Pinto
ECBio, Investigação e Desenvolvimento em Biotecnologia S.A., R. Henrique Paiva Couceiro, 27, 2700-4511, Amadora, Portugal.
Pedro E
PE
Cruz
ECBio, Investigação e Desenvolvimento em Biotecnologia S.A., R. Henrique Paiva Couceiro, 27, 2700-4511, Amadora, Portugal.
Helder J
HJ
Cruz
Centro Cardiovascular da Universidade de Lisboa, Av. Prof Egas Moniz, 1649-028, Lisbon, Portugal. sconstantino@medicina.ulisboa.pt.|Faculdade de Medicina da Universidade de Lisboa, Av. Prof Egas Moniz, 1649-028, Lisbon, Portugal. sconstantino@medicina.ulisboa.pt.
Susana Constantino Rosa
SC
Santos
Angiogenesis; Arteriogenesis; Critical limb ischemia; Endothelial cells; Hindlimb ischemia; Mesenchymal stem cells; UCX®
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