Limits...
Enhancement of Ischemic Wound Healing by Spheroid Grafting of Human Adipose-Derived Stem Cells Treated with Low-Level Light Irradiation.

Park IS, Chung PS, Ahn JC - PLoS ONE (2015)

Bottom Line: PBS, human adipose tissue-derived stromal cells, and the ASC spheroid were transplanted into a wound bed in athymic mice to evaluate the therapeutic effects of the ASC spheroid in vivo.The ASC spheroid transplanted into the wound bed differentiated into endothelial cells and remained differentiated.The density of vascular formations increased as a result of the angiogenic factors released by the wound bed and enhanced tissue regeneration at the lesion site.

View Article: PubMed Central - PubMed

Affiliation: Beckman Laser Institute Korea, Dankook University, 119 Dandae-ro, Cheonan, Chungnam, 330-714, Korea.

ABSTRACT
We investigated whether low-level light irradiation prior to transplantation of adipose-derived stromal cell (ASC) spheroids in an animal skin wound model stimulated angiogenesis and tissue regeneration to improve functional recovery of skin tissue. The spheroid, composed of hASCs, was irradiated with low-level light and expressed angiogenic factors, including vascular endothelial growth factor (VEGF), basic fibroblast growth factor (FGF), and hepatocyte growth factor (HGF). Immunochemical staining analysis revealed that the spheroid of the hASCs was CD31+, KDR+, and CD34+. On the other hand, monolayer-cultured hASCs were negative for these markers. PBS, human adipose tissue-derived stromal cells, and the ASC spheroid were transplanted into a wound bed in athymic mice to evaluate the therapeutic effects of the ASC spheroid in vivo. The ASC spheroid transplanted into the wound bed differentiated into endothelial cells and remained differentiated. The density of vascular formations increased as a result of the angiogenic factors released by the wound bed and enhanced tissue regeneration at the lesion site. These results indicate that the transplantation of the ASC spheroid significantly improved functional recovery relative to both ASC transplantation and PBS treatment. These findings suggest that transplantation of an ASC spheroid treated with low-level light may be an effective form of stem cell therapy for treatment of a wound bed.

No MeSH data available.


Related in: MedlinePlus

Endothelial phenotyping of L-spheroids.(A) Monitoring cell surface markers via immunofluorescence staining. L-spheroids cultured for 3 days were cryosectioned and stained with anti human CD34, CD31, and KDR antibodies. Scale bar = 500 μm. (B) Flow cytometry analysis.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4465903&req=5

pone.0122776.g002: Endothelial phenotyping of L-spheroids.(A) Monitoring cell surface markers via immunofluorescence staining. L-spheroids cultured for 3 days were cryosectioned and stained with anti human CD34, CD31, and KDR antibodies. Scale bar = 500 μm. (B) Flow cytometry analysis.

Mentions: The endothelial phenotype of the L-spheroid cells was also evaluated via immunofluorescent staining for a variety of endothelial cell surface markers. First, the surface markers of hASCs expanded in DMEM-F12/FBS were examined. The cells expressed CD29 (β1 integrin), CD90 (Thy-1), and CD105 (endoglin), as well as MSC surface antigens, but not CD34, CD31, or KDR (S2B and S2C Fig). This indicated that the cells used in the study included a large population of hASCs without endothelial lineage cell contamination. Immunofluorescence staining also revealed that L-spheroids expressed a variety of EC surface markers, including CD34, CD31, and KDR (VEGF receptor) (Fig 1B). The cell population of the L-spheroids was further characterized via flow cytometry. In a monolayer culture with LLLI, EC markers (CD34, CD31, and KDR) were detected in less than 1% of cells. Conversely, L-spheroids were composed of a population of cells positive for CD34, CD31, and KDR (Fig 2B).


Enhancement of Ischemic Wound Healing by Spheroid Grafting of Human Adipose-Derived Stem Cells Treated with Low-Level Light Irradiation.

Park IS, Chung PS, Ahn JC - PLoS ONE (2015)

Endothelial phenotyping of L-spheroids.(A) Monitoring cell surface markers via immunofluorescence staining. L-spheroids cultured for 3 days were cryosectioned and stained with anti human CD34, CD31, and KDR antibodies. Scale bar = 500 μm. (B) Flow cytometry analysis.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4465903&req=5

pone.0122776.g002: Endothelial phenotyping of L-spheroids.(A) Monitoring cell surface markers via immunofluorescence staining. L-spheroids cultured for 3 days were cryosectioned and stained with anti human CD34, CD31, and KDR antibodies. Scale bar = 500 μm. (B) Flow cytometry analysis.
Mentions: The endothelial phenotype of the L-spheroid cells was also evaluated via immunofluorescent staining for a variety of endothelial cell surface markers. First, the surface markers of hASCs expanded in DMEM-F12/FBS were examined. The cells expressed CD29 (β1 integrin), CD90 (Thy-1), and CD105 (endoglin), as well as MSC surface antigens, but not CD34, CD31, or KDR (S2B and S2C Fig). This indicated that the cells used in the study included a large population of hASCs without endothelial lineage cell contamination. Immunofluorescence staining also revealed that L-spheroids expressed a variety of EC surface markers, including CD34, CD31, and KDR (VEGF receptor) (Fig 1B). The cell population of the L-spheroids was further characterized via flow cytometry. In a monolayer culture with LLLI, EC markers (CD34, CD31, and KDR) were detected in less than 1% of cells. Conversely, L-spheroids were composed of a population of cells positive for CD34, CD31, and KDR (Fig 2B).

Bottom Line: PBS, human adipose tissue-derived stromal cells, and the ASC spheroid were transplanted into a wound bed in athymic mice to evaluate the therapeutic effects of the ASC spheroid in vivo.The ASC spheroid transplanted into the wound bed differentiated into endothelial cells and remained differentiated.The density of vascular formations increased as a result of the angiogenic factors released by the wound bed and enhanced tissue regeneration at the lesion site.

View Article: PubMed Central - PubMed

Affiliation: Beckman Laser Institute Korea, Dankook University, 119 Dandae-ro, Cheonan, Chungnam, 330-714, Korea.

ABSTRACT
We investigated whether low-level light irradiation prior to transplantation of adipose-derived stromal cell (ASC) spheroids in an animal skin wound model stimulated angiogenesis and tissue regeneration to improve functional recovery of skin tissue. The spheroid, composed of hASCs, was irradiated with low-level light and expressed angiogenic factors, including vascular endothelial growth factor (VEGF), basic fibroblast growth factor (FGF), and hepatocyte growth factor (HGF). Immunochemical staining analysis revealed that the spheroid of the hASCs was CD31+, KDR+, and CD34+. On the other hand, monolayer-cultured hASCs were negative for these markers. PBS, human adipose tissue-derived stromal cells, and the ASC spheroid were transplanted into a wound bed in athymic mice to evaluate the therapeutic effects of the ASC spheroid in vivo. The ASC spheroid transplanted into the wound bed differentiated into endothelial cells and remained differentiated. The density of vascular formations increased as a result of the angiogenic factors released by the wound bed and enhanced tissue regeneration at the lesion site. These results indicate that the transplantation of the ASC spheroid significantly improved functional recovery relative to both ASC transplantation and PBS treatment. These findings suggest that transplantation of an ASC spheroid treated with low-level light may be an effective form of stem cell therapy for treatment of a wound bed.

No MeSH data available.


Related in: MedlinePlus