Limits...
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

Localisation of EPCs following in vitro transplantation. Transplantation into EC tubules performed using 50% (4 × 104) or 10% (8 × 103) Qdot-labelled EPCs at 5 h. EPC localisation in (A) 50% and (B) 10% transplantation assays, showing bright field, fluorescence and merged images from 6-14 h at 2 h intervals. Scale bars = 500 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3108917&req=5

Figure 7: Localisation of EPCs following in vitro transplantation. Transplantation into EC tubules performed using 50% (4 × 104) or 10% (8 × 103) Qdot-labelled EPCs at 5 h. EPC localisation in (A) 50% and (B) 10% transplantation assays, showing bright field, fluorescence and merged images from 6-14 h at 2 h intervals. Scale bars = 500 μm.

Mentions: By labeling EPCs with fluorescent Qdots prior to transplantation, their localisation into existing EC tubules could be determined. With 50% transplantation, Qdots were detected in high abundance throughout the assay at all time-points (Figure 7a). Whilst Qdots were not detected ubiquitously throughout the existing EC network, between 6 h and 14 h tubule structures were observed, composed entirely of Qdot-labelled EPCs. Following 10% transplantation, Qdots were randomly distributed within the existing EC tubule network (Figure 7b).


Angiogenic potential of endothelial progenitor cells and embryonic stem cells.

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

Localisation of EPCs following in vitro transplantation. Transplantation into EC tubules performed using 50% (4 × 104) or 10% (8 × 103) Qdot-labelled EPCs at 5 h. EPC localisation in (A) 50% and (B) 10% transplantation assays, showing bright field, fluorescence and merged images from 6-14 h at 2 h intervals. Scale bars = 500 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 7: Localisation of EPCs following in vitro transplantation. Transplantation into EC tubules performed using 50% (4 × 104) or 10% (8 × 103) Qdot-labelled EPCs at 5 h. EPC localisation in (A) 50% and (B) 10% transplantation assays, showing bright field, fluorescence and merged images from 6-14 h at 2 h intervals. Scale bars = 500 μm.
Mentions: By labeling EPCs with fluorescent Qdots prior to transplantation, their localisation into existing EC tubules could be determined. With 50% transplantation, Qdots were detected in high abundance throughout the assay at all time-points (Figure 7a). Whilst Qdots were not detected ubiquitously throughout the existing EC network, between 6 h and 14 h tubule structures were observed, composed entirely of Qdot-labelled EPCs. Following 10% transplantation, Qdots were randomly distributed within the existing EC tubule network (Figure 7b).

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