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Mesenchymal stromal cells enhance the engraftment of hematopoietic stem cells in an autologous mouse transplantation model.

Fernández-García M, Yañez RM, Sánchez-Domínguez R, Hernando-Rodriguez M, Peces-Barba M, Herrera G, O'Connor JE, Segovia JC, Bueren JA, Lamana ML - Stem Cell Res Ther (2015)

Bottom Line: This effect was Ad-MSC dose-dependent and associated with an increased homing of transplanted HSCs in recipients' bone marrow.In vivo and in vitro experiments also indicate that the Ad-MSC effects observed in this autologous transplant model are not due to paracrine effects but rather are related to Ad-MSC and HSC interactions, allowing us to propose that Ad-MSCs may act as HSC carriers, facilitating the migration and homing of the HSCs to recipient bone marrow niches.Our results demonstrate that Ad-MSCs facilitate the engraftment of purified HSCs in an autologous mouse transplantation model, opening new perspectives in the application of Ad-MSCs in autologous transplants, including HSC gene therapy.

View Article: PubMed Central - PubMed

Affiliation: Hematopoietic Innovative Therapies Division. Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas (CIEMAT) and Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBER-ER), Madrid, Spain. maria.fernandez@ciemat.es.

ABSTRACT

Introduction: Studies have proposed that mesenchymal stem cells (MSCs) improve the hematopoietic engraftment in allogeneic or xenogeneic transplants and this is probably due to the MSCs' immunosuppressive properties. Our study aimed to discern, for the first time, whether MSC infusion could facilitate the engraftment of hematopoietic stem cells (HSCs) in autologous transplantations models, where no immune rejection of donor HSCs is expected.

Methods: Recipient mice (CD45.2) mice, conditioned with moderate doses of radiation (5-7 Gy), were transplanted with low numbers of HSCs (CD45.1/CD45.2) either as a sole population or co-infused with increasing numbers of adipose-derived-MSCs (Ad-MSCs). The influence of Ad-MSC infusion on the short-term and long-term engraftment of donor HSCs was investigated. Additionally, homing assays and studies related with the administration route and with the Ad-MSC/HSC interaction were conducted.

Results: Our data show that the co-infusion of Ad-MSCs with low numbers of purified HSCs significantly improves the short-term and long-term hematopoietic reconstitution of recipients conditioned with moderate irradiation doses. This effect was Ad-MSC dose-dependent and associated with an increased homing of transplanted HSCs in recipients' bone marrow. In vivo and in vitro experiments also indicate that the Ad-MSC effects observed in this autologous transplant model are not due to paracrine effects but rather are related to Ad-MSC and HSC interactions, allowing us to propose that Ad-MSCs may act as HSC carriers, facilitating the migration and homing of the HSCs to recipient bone marrow niches.

Conclusion: Our results demonstrate that Ad-MSCs facilitate the engraftment of purified HSCs in an autologous mouse transplantation model, opening new perspectives in the application of Ad-MSCs in autologous transplants, including HSC gene therapy.

No MeSH data available.


Flow cytometry analysis of the rapid in vitro interaction of HSCs with Ad-MSCs. a Representative flow cytometry duplet analysis of 6×105 CFSE-stained Ad-MSCs and 1500 DiD-stained LSK cells, 10 minutes after co-culture. b Percentage of DiD+LSK cells forming duplets with CFSE+ Ad-MSCs detected in the different assays. c Multi-spectral images of PE+ LSK cells and FITC+ Ad-MSCs. Individual cells and three representative duplets are shown by a row of images that include side scatter signal (blue), FITC signal (green), PE signal (orange), and FITC and PE signals together (merged column). Bright-field (BF) of each cell sample is also represented. Ad-MSC adipose tissue-derived mesenchymal stem cell, CFSE carboxyfluorescein succinimidyl ester, DAPI 4',6-diamidino-2-phenylindole, FITC fluorescein isothiocyanate, FSC forward scatter, HSC hematopoietic stem cell, LSK lineage− Sca-1+ cKit+, PE phycoerythrin, SSC side scatter
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Fig8: Flow cytometry analysis of the rapid in vitro interaction of HSCs with Ad-MSCs. a Representative flow cytometry duplet analysis of 6×105 CFSE-stained Ad-MSCs and 1500 DiD-stained LSK cells, 10 minutes after co-culture. b Percentage of DiD+LSK cells forming duplets with CFSE+ Ad-MSCs detected in the different assays. c Multi-spectral images of PE+ LSK cells and FITC+ Ad-MSCs. Individual cells and three representative duplets are shown by a row of images that include side scatter signal (blue), FITC signal (green), PE signal (orange), and FITC and PE signals together (merged column). Bright-field (BF) of each cell sample is also represented. Ad-MSC adipose tissue-derived mesenchymal stem cell, CFSE carboxyfluorescein succinimidyl ester, DAPI 4',6-diamidino-2-phenylindole, FITC fluorescein isothiocyanate, FSC forward scatter, HSC hematopoietic stem cell, LSK lineage− Sca-1+ cKit+, PE phycoerythrin, SSC side scatter

Mentions: This hypothesis was further supported by the finding that after a short incubation (10 minutes) of DiD+ LSK cells with CSFE+ Ad-MSCs, 83.69 ± 2 % of the LSK cells formed DiD+/CFSE+ cell duplets, showing the close interaction between both cell types (Fig. 8a, b). In similar experiments, co-incubated PE+ LSK and fluorescein isothiocyanate-positive (FITC+) Ad-MSCs were analyzed by image stream flow cytometry methods to examine the direct interaction of the two cell populations. In Fig. 8c, the intimate association of LSK and Ad-MSCs is shown. These final observations suggest that Ad-MSCs may act as LSK carriers in vivo, thus facilitating the migration and homing of the HSC and hematopoietic progenitor cells into the BM niche during the early stages after transplantation.Fig. 8


Mesenchymal stromal cells enhance the engraftment of hematopoietic stem cells in an autologous mouse transplantation model.

Fernández-García M, Yañez RM, Sánchez-Domínguez R, Hernando-Rodriguez M, Peces-Barba M, Herrera G, O'Connor JE, Segovia JC, Bueren JA, Lamana ML - Stem Cell Res Ther (2015)

Flow cytometry analysis of the rapid in vitro interaction of HSCs with Ad-MSCs. a Representative flow cytometry duplet analysis of 6×105 CFSE-stained Ad-MSCs and 1500 DiD-stained LSK cells, 10 minutes after co-culture. b Percentage of DiD+LSK cells forming duplets with CFSE+ Ad-MSCs detected in the different assays. c Multi-spectral images of PE+ LSK cells and FITC+ Ad-MSCs. Individual cells and three representative duplets are shown by a row of images that include side scatter signal (blue), FITC signal (green), PE signal (orange), and FITC and PE signals together (merged column). Bright-field (BF) of each cell sample is also represented. Ad-MSC adipose tissue-derived mesenchymal stem cell, CFSE carboxyfluorescein succinimidyl ester, DAPI 4',6-diamidino-2-phenylindole, FITC fluorescein isothiocyanate, FSC forward scatter, HSC hematopoietic stem cell, LSK lineage− Sca-1+ cKit+, PE phycoerythrin, SSC side scatter
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4562358&req=5

Fig8: Flow cytometry analysis of the rapid in vitro interaction of HSCs with Ad-MSCs. a Representative flow cytometry duplet analysis of 6×105 CFSE-stained Ad-MSCs and 1500 DiD-stained LSK cells, 10 minutes after co-culture. b Percentage of DiD+LSK cells forming duplets with CFSE+ Ad-MSCs detected in the different assays. c Multi-spectral images of PE+ LSK cells and FITC+ Ad-MSCs. Individual cells and three representative duplets are shown by a row of images that include side scatter signal (blue), FITC signal (green), PE signal (orange), and FITC and PE signals together (merged column). Bright-field (BF) of each cell sample is also represented. Ad-MSC adipose tissue-derived mesenchymal stem cell, CFSE carboxyfluorescein succinimidyl ester, DAPI 4',6-diamidino-2-phenylindole, FITC fluorescein isothiocyanate, FSC forward scatter, HSC hematopoietic stem cell, LSK lineage− Sca-1+ cKit+, PE phycoerythrin, SSC side scatter
Mentions: This hypothesis was further supported by the finding that after a short incubation (10 minutes) of DiD+ LSK cells with CSFE+ Ad-MSCs, 83.69 ± 2 % of the LSK cells formed DiD+/CFSE+ cell duplets, showing the close interaction between both cell types (Fig. 8a, b). In similar experiments, co-incubated PE+ LSK and fluorescein isothiocyanate-positive (FITC+) Ad-MSCs were analyzed by image stream flow cytometry methods to examine the direct interaction of the two cell populations. In Fig. 8c, the intimate association of LSK and Ad-MSCs is shown. These final observations suggest that Ad-MSCs may act as LSK carriers in vivo, thus facilitating the migration and homing of the HSC and hematopoietic progenitor cells into the BM niche during the early stages after transplantation.Fig. 8

Bottom Line: This effect was Ad-MSC dose-dependent and associated with an increased homing of transplanted HSCs in recipients' bone marrow.In vivo and in vitro experiments also indicate that the Ad-MSC effects observed in this autologous transplant model are not due to paracrine effects but rather are related to Ad-MSC and HSC interactions, allowing us to propose that Ad-MSCs may act as HSC carriers, facilitating the migration and homing of the HSCs to recipient bone marrow niches.Our results demonstrate that Ad-MSCs facilitate the engraftment of purified HSCs in an autologous mouse transplantation model, opening new perspectives in the application of Ad-MSCs in autologous transplants, including HSC gene therapy.

View Article: PubMed Central - PubMed

Affiliation: Hematopoietic Innovative Therapies Division. Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas (CIEMAT) and Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBER-ER), Madrid, Spain. maria.fernandez@ciemat.es.

ABSTRACT

Introduction: Studies have proposed that mesenchymal stem cells (MSCs) improve the hematopoietic engraftment in allogeneic or xenogeneic transplants and this is probably due to the MSCs' immunosuppressive properties. Our study aimed to discern, for the first time, whether MSC infusion could facilitate the engraftment of hematopoietic stem cells (HSCs) in autologous transplantations models, where no immune rejection of donor HSCs is expected.

Methods: Recipient mice (CD45.2) mice, conditioned with moderate doses of radiation (5-7 Gy), were transplanted with low numbers of HSCs (CD45.1/CD45.2) either as a sole population or co-infused with increasing numbers of adipose-derived-MSCs (Ad-MSCs). The influence of Ad-MSC infusion on the short-term and long-term engraftment of donor HSCs was investigated. Additionally, homing assays and studies related with the administration route and with the Ad-MSC/HSC interaction were conducted.

Results: Our data show that the co-infusion of Ad-MSCs with low numbers of purified HSCs significantly improves the short-term and long-term hematopoietic reconstitution of recipients conditioned with moderate irradiation doses. This effect was Ad-MSC dose-dependent and associated with an increased homing of transplanted HSCs in recipients' bone marrow. In vivo and in vitro experiments also indicate that the Ad-MSC effects observed in this autologous transplant model are not due to paracrine effects but rather are related to Ad-MSC and HSC interactions, allowing us to propose that Ad-MSCs may act as HSC carriers, facilitating the migration and homing of the HSCs to recipient bone marrow niches.

Conclusion: Our results demonstrate that Ad-MSCs facilitate the engraftment of purified HSCs in an autologous mouse transplantation model, opening new perspectives in the application of Ad-MSCs in autologous transplants, including HSC gene therapy.

No MeSH data available.