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Adenoviral transduction of mesenchymal stem cells: in vitro responses and in vivo immune responses after cell transplantation.

Treacy O, Ryan AE, Heinzl T, O'Flynn L, Cregg M, Wilk M, Odoardi F, Lohan P, O'Brien T, Nosov M, Ritter T - PLoS ONE (2012)

Bottom Line: We found that Ad-transduction of MSCs does not lead to up-regulation of major histocompatibility complex class I and II and co-stimulatory molecules CD80 and CD86.In vivo injection of Ad-transduced MSCs did not change the frequency of various immune cell populations (antigen presenting cells, T helper and cytotoxic T cells, natural killer and natural killer T cells) neither in the blood nor in tissues.Our results indicate that Ad-modification has no major influence on the immunological properties of MSCs and therefore can be considered as a suitable gene vector for therapeutic applications of MSCs.

View Article: PubMed Central - PubMed

Affiliation: College of Medicine, Nursing and Health Sciences, School of Medicine, Regenerative Medicine Institute, National University of Ireland, Galway, Ireland.

ABSTRACT
Adult mesenchymal stem cells (MSCs) are non-hematopoietic cells with multi-lineage potential which makes them attractive targets for regenerative medicine applications. However, to date, therapeutic success of MSC-therapy is limited and the genetic modification of MSCs using viral vectors is one option to improve their therapeutic potential. Ex-vivo genetic modification of MSCs using recombinant adenovirus (Ad) could be promising to reduce undesired immune responses as Ad will be removed before cell/tissue transplantation. In this regard, we investigated whether Ad-modification of MSCs alters their immunological properties in vitro and in vivo. We found that Ad-transduction of MSCs does not lead to up-regulation of major histocompatibility complex class I and II and co-stimulatory molecules CD80 and CD86. Moreover, Ad-transduction caused no significant changes in terms of pro-inflammatory cytokine expression, chemokine and chemokine receptor and Toll-like receptor expression. In addition, Ad-modification of MSCs had no affect on their ability to suppress T cell proliferation in vitro. In vivo injection of Ad-transduced MSCs did not change the frequency of various immune cell populations (antigen presenting cells, T helper and cytotoxic T cells, natural killer and natural killer T cells) neither in the blood nor in tissues. Our results indicate that Ad-modification has no major influence on the immunological properties of MSCs and therefore can be considered as a suitable gene vector for therapeutic applications of MSCs.

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Related in: MedlinePlus

Cell surface characterization and differentiation potential of MSCs.(A) Cell surface expression of various MSC markers were detected by staining with specific monoclonal antibodies and analysed by flow cytometry. MSCs are CD29, CD73, CD90 and CD44H positive, and CD45RA, CD71 low or negative. Shown are FACS histograms of CD MSCs stained with antibodies against surface markers as indicated or with appropriate isotype controls in triplicates. MSCs were tested for their potential to differentiate into (B) adipocytes and (C) osteocytes by incubating the cells with specific formulations. Results shown are representative data of 3 separate isolations.
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pone-0042662-g001: Cell surface characterization and differentiation potential of MSCs.(A) Cell surface expression of various MSC markers were detected by staining with specific monoclonal antibodies and analysed by flow cytometry. MSCs are CD29, CD73, CD90 and CD44H positive, and CD45RA, CD71 low or negative. Shown are FACS histograms of CD MSCs stained with antibodies against surface markers as indicated or with appropriate isotype controls in triplicates. MSCs were tested for their potential to differentiate into (B) adipocytes and (C) osteocytes by incubating the cells with specific formulations. Results shown are representative data of 3 separate isolations.

Mentions: Rat MSCs were isolated from bone marrow (BM) of CD rats, cultured and subsequently characterized for the expression of relevant cell surface markers and their potential to differentiate into adipocytes and osteocytes. Flow cytometric analysis showed that MSCs express the cell surface markers CD29, CD73, CD90, and CD44H and lowly express or are negative for CD45RA and CD71 (Figure 1A). MSCs can differentiate along the adipogenic and osteogenic lineages as measured by Oil Red O quantification and calcium concentration (Figure 1B and C, respectively). In summary our results show that the cells isolated from BM of rats are truly MSCs which are being used in all subsequent experiments.


Adenoviral transduction of mesenchymal stem cells: in vitro responses and in vivo immune responses after cell transplantation.

Treacy O, Ryan AE, Heinzl T, O'Flynn L, Cregg M, Wilk M, Odoardi F, Lohan P, O'Brien T, Nosov M, Ritter T - PLoS ONE (2012)

Cell surface characterization and differentiation potential of MSCs.(A) Cell surface expression of various MSC markers were detected by staining with specific monoclonal antibodies and analysed by flow cytometry. MSCs are CD29, CD73, CD90 and CD44H positive, and CD45RA, CD71 low or negative. Shown are FACS histograms of CD MSCs stained with antibodies against surface markers as indicated or with appropriate isotype controls in triplicates. MSCs were tested for their potential to differentiate into (B) adipocytes and (C) osteocytes by incubating the cells with specific formulations. Results shown are representative data of 3 separate isolations.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3412834&req=5

pone-0042662-g001: Cell surface characterization and differentiation potential of MSCs.(A) Cell surface expression of various MSC markers were detected by staining with specific monoclonal antibodies and analysed by flow cytometry. MSCs are CD29, CD73, CD90 and CD44H positive, and CD45RA, CD71 low or negative. Shown are FACS histograms of CD MSCs stained with antibodies against surface markers as indicated or with appropriate isotype controls in triplicates. MSCs were tested for their potential to differentiate into (B) adipocytes and (C) osteocytes by incubating the cells with specific formulations. Results shown are representative data of 3 separate isolations.
Mentions: Rat MSCs were isolated from bone marrow (BM) of CD rats, cultured and subsequently characterized for the expression of relevant cell surface markers and their potential to differentiate into adipocytes and osteocytes. Flow cytometric analysis showed that MSCs express the cell surface markers CD29, CD73, CD90, and CD44H and lowly express or are negative for CD45RA and CD71 (Figure 1A). MSCs can differentiate along the adipogenic and osteogenic lineages as measured by Oil Red O quantification and calcium concentration (Figure 1B and C, respectively). In summary our results show that the cells isolated from BM of rats are truly MSCs which are being used in all subsequent experiments.

Bottom Line: We found that Ad-transduction of MSCs does not lead to up-regulation of major histocompatibility complex class I and II and co-stimulatory molecules CD80 and CD86.In vivo injection of Ad-transduced MSCs did not change the frequency of various immune cell populations (antigen presenting cells, T helper and cytotoxic T cells, natural killer and natural killer T cells) neither in the blood nor in tissues.Our results indicate that Ad-modification has no major influence on the immunological properties of MSCs and therefore can be considered as a suitable gene vector for therapeutic applications of MSCs.

View Article: PubMed Central - PubMed

Affiliation: College of Medicine, Nursing and Health Sciences, School of Medicine, Regenerative Medicine Institute, National University of Ireland, Galway, Ireland.

ABSTRACT
Adult mesenchymal stem cells (MSCs) are non-hematopoietic cells with multi-lineage potential which makes them attractive targets for regenerative medicine applications. However, to date, therapeutic success of MSC-therapy is limited and the genetic modification of MSCs using viral vectors is one option to improve their therapeutic potential. Ex-vivo genetic modification of MSCs using recombinant adenovirus (Ad) could be promising to reduce undesired immune responses as Ad will be removed before cell/tissue transplantation. In this regard, we investigated whether Ad-modification of MSCs alters their immunological properties in vitro and in vivo. We found that Ad-transduction of MSCs does not lead to up-regulation of major histocompatibility complex class I and II and co-stimulatory molecules CD80 and CD86. Moreover, Ad-transduction caused no significant changes in terms of pro-inflammatory cytokine expression, chemokine and chemokine receptor and Toll-like receptor expression. In addition, Ad-modification of MSCs had no affect on their ability to suppress T cell proliferation in vitro. In vivo injection of Ad-transduced MSCs did not change the frequency of various immune cell populations (antigen presenting cells, T helper and cytotoxic T cells, natural killer and natural killer T cells) neither in the blood nor in tissues. Our results indicate that Ad-modification has no major influence on the immunological properties of MSCs and therefore can be considered as a suitable gene vector for therapeutic applications of MSCs.

Show MeSH
Related in: MedlinePlus