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Increased expression of pigment epithelium-derived factor in aged mesenchymal stem cells impairs their therapeutic efficacy for attenuating myocardial infarction injury.

Liang H, Hou H, Yi W, Yang G, Gu C, Lau WB, Gao E, Ma X, Lu Z, Wei X, Pei J, Yi D - Eur. Heart J. (2011)

Bottom Line: Knocking down PEDF expression in old MSCs improved MSC therapeutic efficacy, and induced a cellular profile similar to young MSCs administration.Furthermore, the impaired therapeutic ability of aged MSCs is predominantly caused by increased PEDF secretion.These findings indicate PEDF as a promising novel genetic modification target for improving aged MSC therapeutic efficacy.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiovascular Surgery, Institute of Cardiovascular Disease of Chinese PLA, Xijing Hospital, the Fourth Military Medical University, No.127, West Changle Road, Xi'an, Shaanxi Province 710032, China.

ABSTRACT

Aims: Mesenchymal stem cells (MSCs) can ameliorate myocardial infarction (MI) injury. However, older-donor MSCs seem less efficacious than those from younger donors, and the contributing underlying mechanisms remain unknown. Here, we determine how age-related expression of pigment epithelium-derived factor (PEDF) affects MSC therapeutic efficacy for MI.

Methods and results: Reverse transcriptase-polymerized chain reaction  and enzyme-linked immunosorbent assay analyses revealed dramatically increased PEDF expression in MSCs from old mice compared to young mice. Morphological and functional experiments demonstrated significantly impaired old MSC therapeutic efficacy compared with young MSCs in treatment of mice subjected to MI. Immunofluorescent staining demonstrated that administration of old MSCs compared with young MSCs resulted in an infarct region containing fewer endothelial cells, vascular smooth muscle cells, and macrophages, but more fibroblasts. Pigment epithelium-derived factor overexpression in young MSCs impaired the beneficial effects against MI injury, and induced cellular profile changes in the infarct region similar to administration of old MSCs. Knocking down PEDF expression in old MSCs improved MSC therapeutic efficacy, and induced a cellular profile similar to young MSCs administration. Studies in vitro showed that PEDF secreted by MSCs regulated the proliferation and migration of cardiac fibroblasts.

Conclusions: This is the first evidence that paracrine factor PEDF plays critical role in the regulatory effects of MSCs against MI injury. Furthermore, the impaired therapeutic ability of aged MSCs is predominantly caused by increased PEDF secretion. These findings indicate PEDF as a promising novel genetic modification target for improving aged MSC therapeutic efficacy.

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Characterization of mesenchymal stem cells and expression of pigment epithelium-derived factor. (A) Mesenchymal stem cells (passage 3) from both young and older donor mice display fibroblast-like morphology in culture. In vitro adipocyte differentiation was confirmed by Oil Red O staining (arrows show accumulation of lipid droplets in vacuoles). In vitro osteogenic differentiation was demonstrated by alizarin red staining, demonstrating Ca2+ deposition (arrows). (B) Reverse transcriptase–polymerized chain reaction results demonstrated pigment epithelium-derived factor mRNA levels in older mesenchymal stem cells and young mesenchymal stem cells under both normoxic/hypoxic conditions (n= 5 mice/group). (C ) Enzyme-linked immuno sorbent assay experiment demonstrated pigment epithelium-derived factor protein levels in older mesenchymal stem cells and young mesenchymal stem cells culture supernatants (n= 5 mice/group) Data expressed as means ± SEM. *P< 0.01, **P< 0.001.
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EHR131F1: Characterization of mesenchymal stem cells and expression of pigment epithelium-derived factor. (A) Mesenchymal stem cells (passage 3) from both young and older donor mice display fibroblast-like morphology in culture. In vitro adipocyte differentiation was confirmed by Oil Red O staining (arrows show accumulation of lipid droplets in vacuoles). In vitro osteogenic differentiation was demonstrated by alizarin red staining, demonstrating Ca2+ deposition (arrows). (B) Reverse transcriptase–polymerized chain reaction results demonstrated pigment epithelium-derived factor mRNA levels in older mesenchymal stem cells and young mesenchymal stem cells under both normoxic/hypoxic conditions (n= 5 mice/group). (C ) Enzyme-linked immuno sorbent assay experiment demonstrated pigment epithelium-derived factor protein levels in older mesenchymal stem cells and young mesenchymal stem cells culture supernatants (n= 5 mice/group) Data expressed as means ± SEM. *P< 0.01, **P< 0.001.

Mentions: After three passages, MSC pluripotency was confirmed by capacity to differentiate into adipogenic and osteogenic lineages in vitro (Figure 1A). By flow cytometry analysis, cultured MSCs expressed CD29, CD44, CD73, CD90, and CD105, and were devoid of haematopoietic markers CD11b, CD34, and CD45 (Supplementary material online, Table S1). Notably, RT–PCR (Figure 1B) and ELISA (Figure 1C) revealed that, in comparison to young MSCs, significantly greater PEDF expression (for both mRNA and protein levels) was found in older MSCs under both normoxic and hypoxic conditions.Figure 1


Increased expression of pigment epithelium-derived factor in aged mesenchymal stem cells impairs their therapeutic efficacy for attenuating myocardial infarction injury.

Liang H, Hou H, Yi W, Yang G, Gu C, Lau WB, Gao E, Ma X, Lu Z, Wei X, Pei J, Yi D - Eur. Heart J. (2011)

Characterization of mesenchymal stem cells and expression of pigment epithelium-derived factor. (A) Mesenchymal stem cells (passage 3) from both young and older donor mice display fibroblast-like morphology in culture. In vitro adipocyte differentiation was confirmed by Oil Red O staining (arrows show accumulation of lipid droplets in vacuoles). In vitro osteogenic differentiation was demonstrated by alizarin red staining, demonstrating Ca2+ deposition (arrows). (B) Reverse transcriptase–polymerized chain reaction results demonstrated pigment epithelium-derived factor mRNA levels in older mesenchymal stem cells and young mesenchymal stem cells under both normoxic/hypoxic conditions (n= 5 mice/group). (C ) Enzyme-linked immuno sorbent assay experiment demonstrated pigment epithelium-derived factor protein levels in older mesenchymal stem cells and young mesenchymal stem cells culture supernatants (n= 5 mice/group) Data expressed as means ± SEM. *P< 0.01, **P< 0.001.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3675387&req=5

EHR131F1: Characterization of mesenchymal stem cells and expression of pigment epithelium-derived factor. (A) Mesenchymal stem cells (passage 3) from both young and older donor mice display fibroblast-like morphology in culture. In vitro adipocyte differentiation was confirmed by Oil Red O staining (arrows show accumulation of lipid droplets in vacuoles). In vitro osteogenic differentiation was demonstrated by alizarin red staining, demonstrating Ca2+ deposition (arrows). (B) Reverse transcriptase–polymerized chain reaction results demonstrated pigment epithelium-derived factor mRNA levels in older mesenchymal stem cells and young mesenchymal stem cells under both normoxic/hypoxic conditions (n= 5 mice/group). (C ) Enzyme-linked immuno sorbent assay experiment demonstrated pigment epithelium-derived factor protein levels in older mesenchymal stem cells and young mesenchymal stem cells culture supernatants (n= 5 mice/group) Data expressed as means ± SEM. *P< 0.01, **P< 0.001.
Mentions: After three passages, MSC pluripotency was confirmed by capacity to differentiate into adipogenic and osteogenic lineages in vitro (Figure 1A). By flow cytometry analysis, cultured MSCs expressed CD29, CD44, CD73, CD90, and CD105, and were devoid of haematopoietic markers CD11b, CD34, and CD45 (Supplementary material online, Table S1). Notably, RT–PCR (Figure 1B) and ELISA (Figure 1C) revealed that, in comparison to young MSCs, significantly greater PEDF expression (for both mRNA and protein levels) was found in older MSCs under both normoxic and hypoxic conditions.Figure 1

Bottom Line: Knocking down PEDF expression in old MSCs improved MSC therapeutic efficacy, and induced a cellular profile similar to young MSCs administration.Furthermore, the impaired therapeutic ability of aged MSCs is predominantly caused by increased PEDF secretion.These findings indicate PEDF as a promising novel genetic modification target for improving aged MSC therapeutic efficacy.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiovascular Surgery, Institute of Cardiovascular Disease of Chinese PLA, Xijing Hospital, the Fourth Military Medical University, No.127, West Changle Road, Xi'an, Shaanxi Province 710032, China.

ABSTRACT

Aims: Mesenchymal stem cells (MSCs) can ameliorate myocardial infarction (MI) injury. However, older-donor MSCs seem less efficacious than those from younger donors, and the contributing underlying mechanisms remain unknown. Here, we determine how age-related expression of pigment epithelium-derived factor (PEDF) affects MSC therapeutic efficacy for MI.

Methods and results: Reverse transcriptase-polymerized chain reaction  and enzyme-linked immunosorbent assay analyses revealed dramatically increased PEDF expression in MSCs from old mice compared to young mice. Morphological and functional experiments demonstrated significantly impaired old MSC therapeutic efficacy compared with young MSCs in treatment of mice subjected to MI. Immunofluorescent staining demonstrated that administration of old MSCs compared with young MSCs resulted in an infarct region containing fewer endothelial cells, vascular smooth muscle cells, and macrophages, but more fibroblasts. Pigment epithelium-derived factor overexpression in young MSCs impaired the beneficial effects against MI injury, and induced cellular profile changes in the infarct region similar to administration of old MSCs. Knocking down PEDF expression in old MSCs improved MSC therapeutic efficacy, and induced a cellular profile similar to young MSCs administration. Studies in vitro showed that PEDF secreted by MSCs regulated the proliferation and migration of cardiac fibroblasts.

Conclusions: This is the first evidence that paracrine factor PEDF plays critical role in the regulatory effects of MSCs against MI injury. Furthermore, the impaired therapeutic ability of aged MSCs is predominantly caused by increased PEDF secretion. These findings indicate PEDF as a promising novel genetic modification target for improving aged MSC therapeutic efficacy.

Show MeSH
Related in: MedlinePlus