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Endothelial reconstitution by CD34+ progenitors derived from baboon embryonic stem cells.

Shi Q, Schatten G, Hodara V, Simerly C, VandeBerg JL - J. Cell. Mol. Med. (2013)

Bottom Line: The efficiency of generating CD34+ EPCs did not differ significantly from ECGS to EGM-2 culture media, however, angioblasts specified in ECGS medium expressed a higher percentage of CD34+/CXCR4+ cells (3.49 ± 1.32%, n = 3) than those specified in EGM-2 medium (0.49 ± 0.52%, n = 3).After 14 days of ex vivo culture, the grafted cells had attached and integrated to the denuded surface; in addition, they had matured further and expressed terminally differentiated endothelial markers including CD31 and CD146.In conclusion, we have proved that specified CD34+ EPCs are promising therapeutic agents for repairing damaged vasculature.

View Article: PubMed Central - PubMed

Affiliation: Southwest National Primate Research Center, Texas Biomedical Research Institute, San Antonio, TX 78245-0549, USA. qshi@txbiomedgenetics.org

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Endotheliazation of CD34+ EPCs derived from ESCs in ex vivo culture system. Attachment, growth and maturation of CD34+ EPCs on denuded artery surface. Transplanted cells labelled with CMRA attach, grow and mature on the interior surface of denuded blood vessels. The lumen of the vessel is at the upper left. (A) Transplanted cells express CD31 as demonstrated by anti-CD31 conjugated with FITC (green); (B) transplanted cells express CD146 as demonstrated by anti-CD146 conjugated with FITC (green); (C) is a vehicle control. Nuclei are stained blue with DAPI.
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fig03: Endotheliazation of CD34+ EPCs derived from ESCs in ex vivo culture system. Attachment, growth and maturation of CD34+ EPCs on denuded artery surface. Transplanted cells labelled with CMRA attach, grow and mature on the interior surface of denuded blood vessels. The lumen of the vessel is at the upper left. (A) Transplanted cells express CD31 as demonstrated by anti-CD31 conjugated with FITC (green); (B) transplanted cells express CD146 as demonstrated by anti-CD146 conjugated with FITC (green); (C) is a vehicle control. Nuclei are stained blue with DAPI.

Mentions: To test whether specified angioblasts could function as progenitors to execute a reparative role, we examined their ability to re-endothelialize blood vessels whose endothelium was removed by enzymatic digestion. We tested our hypothesis in an ex vivo bioreactor that we constructed. After culturing for approximately 2 weeks, a 0.5–0.7 cm segment was fixed, embedded in OCT and sectioned longitudinally into 10-μm-thick serial sections; whole-mount immunostaining was performed to assess the fate of the transplanted cells. Figure 3 shows cellular images acquired from serial sections of a representative ex vivo sample. Immunofluorescence staining indicated that transplanted CD34+ EPCs not only homed to and recovered the denuded surface but they also matured to fully differentiated cells by expressing CD31 (a) and CD146 (b), which were not expressed in angioblast-derived cells before seeding. This result implies that extracellular matrix of denuded vasculature possesses signalling molecules that direct and promote the transplanted CD34+ EPCs to gain full functional competence.


Endothelial reconstitution by CD34+ progenitors derived from baboon embryonic stem cells.

Shi Q, Schatten G, Hodara V, Simerly C, VandeBerg JL - J. Cell. Mol. Med. (2013)

Endotheliazation of CD34+ EPCs derived from ESCs in ex vivo culture system. Attachment, growth and maturation of CD34+ EPCs on denuded artery surface. Transplanted cells labelled with CMRA attach, grow and mature on the interior surface of denuded blood vessels. The lumen of the vessel is at the upper left. (A) Transplanted cells express CD31 as demonstrated by anti-CD31 conjugated with FITC (green); (B) transplanted cells express CD146 as demonstrated by anti-CD146 conjugated with FITC (green); (C) is a vehicle control. Nuclei are stained blue with DAPI.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3814022&req=5

fig03: Endotheliazation of CD34+ EPCs derived from ESCs in ex vivo culture system. Attachment, growth and maturation of CD34+ EPCs on denuded artery surface. Transplanted cells labelled with CMRA attach, grow and mature on the interior surface of denuded blood vessels. The lumen of the vessel is at the upper left. (A) Transplanted cells express CD31 as demonstrated by anti-CD31 conjugated with FITC (green); (B) transplanted cells express CD146 as demonstrated by anti-CD146 conjugated with FITC (green); (C) is a vehicle control. Nuclei are stained blue with DAPI.
Mentions: To test whether specified angioblasts could function as progenitors to execute a reparative role, we examined their ability to re-endothelialize blood vessels whose endothelium was removed by enzymatic digestion. We tested our hypothesis in an ex vivo bioreactor that we constructed. After culturing for approximately 2 weeks, a 0.5–0.7 cm segment was fixed, embedded in OCT and sectioned longitudinally into 10-μm-thick serial sections; whole-mount immunostaining was performed to assess the fate of the transplanted cells. Figure 3 shows cellular images acquired from serial sections of a representative ex vivo sample. Immunofluorescence staining indicated that transplanted CD34+ EPCs not only homed to and recovered the denuded surface but they also matured to fully differentiated cells by expressing CD31 (a) and CD146 (b), which were not expressed in angioblast-derived cells before seeding. This result implies that extracellular matrix of denuded vasculature possesses signalling molecules that direct and promote the transplanted CD34+ EPCs to gain full functional competence.

Bottom Line: The efficiency of generating CD34+ EPCs did not differ significantly from ECGS to EGM-2 culture media, however, angioblasts specified in ECGS medium expressed a higher percentage of CD34+/CXCR4+ cells (3.49 ± 1.32%, n = 3) than those specified in EGM-2 medium (0.49 ± 0.52%, n = 3).After 14 days of ex vivo culture, the grafted cells had attached and integrated to the denuded surface; in addition, they had matured further and expressed terminally differentiated endothelial markers including CD31 and CD146.In conclusion, we have proved that specified CD34+ EPCs are promising therapeutic agents for repairing damaged vasculature.

View Article: PubMed Central - PubMed

Affiliation: Southwest National Primate Research Center, Texas Biomedical Research Institute, San Antonio, TX 78245-0549, USA. qshi@txbiomedgenetics.org

Show MeSH
Related in: MedlinePlus