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Angiogenic potential of endothelial progenitor cells and embryonic stem cells.

Rae PC, Kelly RD, Egginton S, St John JC - (2011)

Bottom Line: We also described the production of highly angiogenic EPC-comparable cells from pluripotent embryonic stem cells (ESCs) by direct differentiation using EC-conditioned medium (ECCM).ECCM-treated ESC-derived progenitor cells exhibited angiogenic potential, demonstrated by in vitro tubule formation, and endothelial-specific gene expression equivalent to natural EPCs.We concluded the effect of EPCs is cumulative and beneficial, relying on upregulation of the angiogenic activity of transplanted cells combined with an increase in proliferative cell number to produce significant effects upon transplantation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Clinical Sciences Research Institute, Warwick Medical School, University of Warwick, UK. justin.stjohn@monash.edu.

ABSTRACT

Background: Endothelial progenitor cells (EPCs) are implicated in a range of pathological conditions, suggesting a natural therapeutic role for EPCs in angiogenesis. However, current angiogenic therapies involving EPC transplantation are inefficient due to rejection of donor EPCs. One solution is to derive an expanded population of EPCs from stem cells in vitro, to be re-introduced as a therapeutic transplant. To demonstrate the therapeutic potential of EPCs we performed in vitro transplantation of EPCs into endothelial cell (EC) tubules using a gel-based tubule formation assay. We also described the production of highly angiogenic EPC-comparable cells from pluripotent embryonic stem cells (ESCs) by direct differentiation using EC-conditioned medium (ECCM).

Results: The effect on tubule complexity and longevity varied with transplantation quantity: significant effects were observed when tubules were transplanted with a quantity of EPCs equivalent to 50% of the number of ECs originally seeded on to the assay gel but not with 10% EPC transplantation. Gene expression of the endothelial markers VEGFR2, VE-cadherin and CD31, determined by qPCR, also changed dynamically during transplantation. ECCM-treated ESC-derived progenitor cells exhibited angiogenic potential, demonstrated by in vitro tubule formation, and endothelial-specific gene expression equivalent to natural EPCs.

Conclusions: We concluded the effect of EPCs is cumulative and beneficial, relying on upregulation of the angiogenic activity of transplanted cells combined with an increase in proliferative cell number to produce significant effects upon transplantation. Furthermore, EPCs derived from ESCs may be developed for use as a rapidly-expandable alternative for angiogenic transplantation therapy.

No MeSH data available.


Related in: MedlinePlus

Characterization of differentiating ESCs at D7. Tubule formation as (A) node number ± SEM (n = 3) and (B) tubule branch length ± SEM (n = 3; *P < 0.05, **P < 0.01 vs. 2 h). (C) VEGFR2, (D) VE-cadherin and (E) CD31 mean expression in D7 ESCs (n = 3) relative to 60% confluent ECs; *P < 0.05, **P < 0.01 vs. 0 h. Dotted line indicates expression in 60% confluent EPCs.
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Figure 5: Characterization of differentiating ESCs at D7. Tubule formation as (A) node number ± SEM (n = 3) and (B) tubule branch length ± SEM (n = 3; *P < 0.05, **P < 0.01 vs. 2 h). (C) VEGFR2, (D) VE-cadherin and (E) CD31 mean expression in D7 ESCs (n = 3) relative to 60% confluent ECs; *P < 0.05, **P < 0.01 vs. 0 h. Dotted line indicates expression in 60% confluent EPCs.

Mentions: As directed-differentiated ESCs more rapidly adopted patterns of EPC-associated gene expression, we cultured differentiating D7 ESCs on ECMatrix gel to assess their angiogenic potential. The pattern of nodes formed by D7 ESCs was similar to that of both EPCs and ECs (P < 0.05; Figure 5a). To this extent, the number of N1 nodes was maximal at 2 h and decreased throughout the assay. Nevertheless, the number of N2, N3 and N4 nodes increased to their maximum towards the assay's mid point, whilst N5+ nodes were only evident at 8 h. Branch length measurements of D7 ESCs, as for the node data, were similar to EPCs and ECs (P < 0.05; Figure 5b). Mean length increased from 0 h to 6 h, with maximal length observed at 6 h, and no branches were evident by 14 h.


Angiogenic potential of endothelial progenitor cells and embryonic stem cells.

Rae PC, Kelly RD, Egginton S, St John JC - (2011)

Characterization of differentiating ESCs at D7. Tubule formation as (A) node number ± SEM (n = 3) and (B) tubule branch length ± SEM (n = 3; *P < 0.05, **P < 0.01 vs. 2 h). (C) VEGFR2, (D) VE-cadherin and (E) CD31 mean expression in D7 ESCs (n = 3) relative to 60% confluent ECs; *P < 0.05, **P < 0.01 vs. 0 h. Dotted line indicates expression in 60% confluent EPCs.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3108917&req=5

Figure 5: Characterization of differentiating ESCs at D7. Tubule formation as (A) node number ± SEM (n = 3) and (B) tubule branch length ± SEM (n = 3; *P < 0.05, **P < 0.01 vs. 2 h). (C) VEGFR2, (D) VE-cadherin and (E) CD31 mean expression in D7 ESCs (n = 3) relative to 60% confluent ECs; *P < 0.05, **P < 0.01 vs. 0 h. Dotted line indicates expression in 60% confluent EPCs.
Mentions: As directed-differentiated ESCs more rapidly adopted patterns of EPC-associated gene expression, we cultured differentiating D7 ESCs on ECMatrix gel to assess their angiogenic potential. The pattern of nodes formed by D7 ESCs was similar to that of both EPCs and ECs (P < 0.05; Figure 5a). To this extent, the number of N1 nodes was maximal at 2 h and decreased throughout the assay. Nevertheless, the number of N2, N3 and N4 nodes increased to their maximum towards the assay's mid point, whilst N5+ nodes were only evident at 8 h. Branch length measurements of D7 ESCs, as for the node data, were similar to EPCs and ECs (P < 0.05; Figure 5b). Mean length increased from 0 h to 6 h, with maximal length observed at 6 h, and no branches were evident by 14 h.

Bottom Line: We also described the production of highly angiogenic EPC-comparable cells from pluripotent embryonic stem cells (ESCs) by direct differentiation using EC-conditioned medium (ECCM).ECCM-treated ESC-derived progenitor cells exhibited angiogenic potential, demonstrated by in vitro tubule formation, and endothelial-specific gene expression equivalent to natural EPCs.We concluded the effect of EPCs is cumulative and beneficial, relying on upregulation of the angiogenic activity of transplanted cells combined with an increase in proliferative cell number to produce significant effects upon transplantation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Clinical Sciences Research Institute, Warwick Medical School, University of Warwick, UK. justin.stjohn@monash.edu.

ABSTRACT

Background: Endothelial progenitor cells (EPCs) are implicated in a range of pathological conditions, suggesting a natural therapeutic role for EPCs in angiogenesis. However, current angiogenic therapies involving EPC transplantation are inefficient due to rejection of donor EPCs. One solution is to derive an expanded population of EPCs from stem cells in vitro, to be re-introduced as a therapeutic transplant. To demonstrate the therapeutic potential of EPCs we performed in vitro transplantation of EPCs into endothelial cell (EC) tubules using a gel-based tubule formation assay. We also described the production of highly angiogenic EPC-comparable cells from pluripotent embryonic stem cells (ESCs) by direct differentiation using EC-conditioned medium (ECCM).

Results: The effect on tubule complexity and longevity varied with transplantation quantity: significant effects were observed when tubules were transplanted with a quantity of EPCs equivalent to 50% of the number of ECs originally seeded on to the assay gel but not with 10% EPC transplantation. Gene expression of the endothelial markers VEGFR2, VE-cadherin and CD31, determined by qPCR, also changed dynamically during transplantation. ECCM-treated ESC-derived progenitor cells exhibited angiogenic potential, demonstrated by in vitro tubule formation, and endothelial-specific gene expression equivalent to natural EPCs.

Conclusions: We concluded the effect of EPCs is cumulative and beneficial, relying on upregulation of the angiogenic activity of transplanted cells combined with an increase in proliferative cell number to produce significant effects upon transplantation. Furthermore, EPCs derived from ESCs may be developed for use as a rapidly-expandable alternative for angiogenic transplantation therapy.

No MeSH data available.


Related in: MedlinePlus