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Gamma irradiation preserves immunosuppressive potential and inhibits clonogenic capacity of human bone marrow-derived mesenchymal stromal cells.

de Andrade AV, Riewaldt J, Wehner R, Schmitz M, Odendahl M, Bornhäuser M, Tonn T - J. Cell. Mol. Med. (2014)

Bottom Line: We therefore analysed the effect of gamma irradiation on the potency and proliferation of MSCs to elucidate an irradiation dose, which would allow inhibition of MSC proliferation while at the same time preserving their immunosuppressive function.No significant decrease of viable cells was detected, as compared to non-irradiated BM-MSCs.Gamma irradiation does not impair the immunosuppressive capacity of BM-MSCs in vitro and thus might increase the safety of MSC-based cell products in clinical applications.

View Article: PubMed Central - PubMed

Affiliation: Institute for Transfusion Medicine, German Red Cross Blood Donation Service North-East, Dresden, Germany.

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Immunophenotypical profile and differentiation capacity of BM-MSCs. BM-MSCs from different cell lines (n = 4, passage 4–6) were characterized. (A) Different surface markers (CD73, CD90, CD105, CD45, CD31, HLA-DR, HLA-ABC and CD39) were analysed by flow cytometry. Bars represent mean percentage of positive cells ±SEM. (B) Differentiation potential into osteoblasts and adipocytes was assessed by the presence of red-stained calcium precipitates and lipid droplets (indicated by arrows; 40× magnification), as visualized by Alizarin Red and Oil Red O staining, respectively. Non-differentiated control cultures are shown in 10× magnification.
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fig01: Immunophenotypical profile and differentiation capacity of BM-MSCs. BM-MSCs from different cell lines (n = 4, passage 4–6) were characterized. (A) Different surface markers (CD73, CD90, CD105, CD45, CD31, HLA-DR, HLA-ABC and CD39) were analysed by flow cytometry. Bars represent mean percentage of positive cells ±SEM. (B) Differentiation potential into osteoblasts and adipocytes was assessed by the presence of red-stained calcium precipitates and lipid droplets (indicated by arrows; 40× magnification), as visualized by Alizarin Red and Oil Red O staining, respectively. Non-differentiated control cultures are shown in 10× magnification.

Mentions: We initially sought to confirm the identity of BM-MSCs by flow cytometry. As expected, and consistent with criteria defined by the ISCT [36], all cell lines tested (n = 4) exhibited a immunophenotypic profile typical for MSCs. This was reflected in expression of the main MSC surface markers CD73, CD90 and CD105 (mean proportion of cells positive for the respective markers ≥98.9%; Fig.1A). In addition, the isolated cell lines expressed HLA-ABC (mean proportion of positive cells/cell line = 32.2 ± 3.3%; Fig.1A), whereas expression of HLA-DR, CD45 and CD31 was virtually absent (mean proportion of positive cells for each of the respective markers <0.1%; Fig.1A). Furthermore, all cell lines tested were positive for CD39 (35.7 ± 9.1% positive cells; Fig.1A). Together, these data clearly demonstrated that the isolated cells were indeed MSCs.


Gamma irradiation preserves immunosuppressive potential and inhibits clonogenic capacity of human bone marrow-derived mesenchymal stromal cells.

de Andrade AV, Riewaldt J, Wehner R, Schmitz M, Odendahl M, Bornhäuser M, Tonn T - J. Cell. Mol. Med. (2014)

Immunophenotypical profile and differentiation capacity of BM-MSCs. BM-MSCs from different cell lines (n = 4, passage 4–6) were characterized. (A) Different surface markers (CD73, CD90, CD105, CD45, CD31, HLA-DR, HLA-ABC and CD39) were analysed by flow cytometry. Bars represent mean percentage of positive cells ±SEM. (B) Differentiation potential into osteoblasts and adipocytes was assessed by the presence of red-stained calcium precipitates and lipid droplets (indicated by arrows; 40× magnification), as visualized by Alizarin Red and Oil Red O staining, respectively. Non-differentiated control cultures are shown in 10× magnification.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4508157&req=5

fig01: Immunophenotypical profile and differentiation capacity of BM-MSCs. BM-MSCs from different cell lines (n = 4, passage 4–6) were characterized. (A) Different surface markers (CD73, CD90, CD105, CD45, CD31, HLA-DR, HLA-ABC and CD39) were analysed by flow cytometry. Bars represent mean percentage of positive cells ±SEM. (B) Differentiation potential into osteoblasts and adipocytes was assessed by the presence of red-stained calcium precipitates and lipid droplets (indicated by arrows; 40× magnification), as visualized by Alizarin Red and Oil Red O staining, respectively. Non-differentiated control cultures are shown in 10× magnification.
Mentions: We initially sought to confirm the identity of BM-MSCs by flow cytometry. As expected, and consistent with criteria defined by the ISCT [36], all cell lines tested (n = 4) exhibited a immunophenotypic profile typical for MSCs. This was reflected in expression of the main MSC surface markers CD73, CD90 and CD105 (mean proportion of cells positive for the respective markers ≥98.9%; Fig.1A). In addition, the isolated cell lines expressed HLA-ABC (mean proportion of positive cells/cell line = 32.2 ± 3.3%; Fig.1A), whereas expression of HLA-DR, CD45 and CD31 was virtually absent (mean proportion of positive cells for each of the respective markers <0.1%; Fig.1A). Furthermore, all cell lines tested were positive for CD39 (35.7 ± 9.1% positive cells; Fig.1A). Together, these data clearly demonstrated that the isolated cells were indeed MSCs.

Bottom Line: We therefore analysed the effect of gamma irradiation on the potency and proliferation of MSCs to elucidate an irradiation dose, which would allow inhibition of MSC proliferation while at the same time preserving their immunosuppressive function.No significant decrease of viable cells was detected, as compared to non-irradiated BM-MSCs.Gamma irradiation does not impair the immunosuppressive capacity of BM-MSCs in vitro and thus might increase the safety of MSC-based cell products in clinical applications.

View Article: PubMed Central - PubMed

Affiliation: Institute for Transfusion Medicine, German Red Cross Blood Donation Service North-East, Dresden, Germany.

Show MeSH
Related in: MedlinePlus