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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.


Short-term and long-term hematopoietic engraftment of recipient mice co-infused with purified HSCs and Ad-MSCs. a Donor CD45.1+/CD45.2+ cell engraftment detected in CD45.2+ recipients’ peripheral blood. Five gray-irradiated primary recipients were infused 1500 LSK, with (gray bars) or without (white bars) 6×105 Ad-MSC co-infusion. Secondary and tertiary recipients were irradiated with 11 Gy and transplanted with pooled 2×106 BM cells obtained 24 weeks after HSCT from the primary recipients and at 12 weeks after HSCT from the secondary recipients. b Flow cytometry analysis showing the percentage of LSK cells that were of donor CD45.1+/CD45.2+ origin in pooled BM samples from primary recipients (CD45.2+) 24 weeks after transplantation. c Similar analyses conducted in pooled BM samples from secondary recipients (CD45.2+) 12 weeks after transplantation. Bars represent standard error of the mean. *P ≤ 0.05. **P ≤ 0.01. ***P ≤ 0.001. Ad-MSC adipose tissue-derived mesenchymal stem cell, BM bone marrow, HSC hematopoietic stem cell, HSCT hematopoietic stem cell transplant, LSK lineage− Sca-1+ cKit+, ns non-significant difference
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Fig4: Short-term and long-term hematopoietic engraftment of recipient mice co-infused with purified HSCs and Ad-MSCs. a Donor CD45.1+/CD45.2+ cell engraftment detected in CD45.2+ recipients’ peripheral blood. Five gray-irradiated primary recipients were infused 1500 LSK, with (gray bars) or without (white bars) 6×105 Ad-MSC co-infusion. Secondary and tertiary recipients were irradiated with 11 Gy and transplanted with pooled 2×106 BM cells obtained 24 weeks after HSCT from the primary recipients and at 12 weeks after HSCT from the secondary recipients. b Flow cytometry analysis showing the percentage of LSK cells that were of donor CD45.1+/CD45.2+ origin in pooled BM samples from primary recipients (CD45.2+) 24 weeks after transplantation. c Similar analyses conducted in pooled BM samples from secondary recipients (CD45.2+) 12 weeks after transplantation. Bars represent standard error of the mean. *P ≤ 0.05. **P ≤ 0.01. ***P ≤ 0.001. Ad-MSC adipose tissue-derived mesenchymal stem cell, BM bone marrow, HSC hematopoietic stem cell, HSCT hematopoietic stem cell transplant, LSK lineage− Sca-1+ cKit+, ns non-significant difference

Mentions: As observed in previous experiments (Fig. 3), a dose of 6×105 Ad-MSCs significantly increased LSK engraftment in primary recipients during the first weeks after transplant, although at 12 and 24 weeks post-transplantation differences did not reach statistical significance (Fig. 4a). This figure also shows that most of the PB analyses performed in secondary and tertiary recipients revealed improved donor engraftments in the LSK + Ad-MSC group, as compared with the LSK group. Consistent with these observations, the proportion of precursor cells that were of donor origin (percentage of LSK cells that were CD45.1+/CD45.2+) in pooled BM samples that were transplanted in secondary and tertiary recipients was also higher in the LSK + Ad-MSC groups (Fig. 4b, c). These experiments allow us to conclude that the co-infusion of Ad-MSCs improves the engraftment not only of progenitors responsible for the early stages of hematopoietic engraftment but also of long-term repopulating HSCs.Fig. 4


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)

Short-term and long-term hematopoietic engraftment of recipient mice co-infused with purified HSCs and Ad-MSCs. a Donor CD45.1+/CD45.2+ cell engraftment detected in CD45.2+ recipients’ peripheral blood. Five gray-irradiated primary recipients were infused 1500 LSK, with (gray bars) or without (white bars) 6×105 Ad-MSC co-infusion. Secondary and tertiary recipients were irradiated with 11 Gy and transplanted with pooled 2×106 BM cells obtained 24 weeks after HSCT from the primary recipients and at 12 weeks after HSCT from the secondary recipients. b Flow cytometry analysis showing the percentage of LSK cells that were of donor CD45.1+/CD45.2+ origin in pooled BM samples from primary recipients (CD45.2+) 24 weeks after transplantation. c Similar analyses conducted in pooled BM samples from secondary recipients (CD45.2+) 12 weeks after transplantation. Bars represent standard error of the mean. *P ≤ 0.05. **P ≤ 0.01. ***P ≤ 0.001. Ad-MSC adipose tissue-derived mesenchymal stem cell, BM bone marrow, HSC hematopoietic stem cell, HSCT hematopoietic stem cell transplant, LSK lineage− Sca-1+ cKit+, ns non-significant difference
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Fig4: Short-term and long-term hematopoietic engraftment of recipient mice co-infused with purified HSCs and Ad-MSCs. a Donor CD45.1+/CD45.2+ cell engraftment detected in CD45.2+ recipients’ peripheral blood. Five gray-irradiated primary recipients were infused 1500 LSK, with (gray bars) or without (white bars) 6×105 Ad-MSC co-infusion. Secondary and tertiary recipients were irradiated with 11 Gy and transplanted with pooled 2×106 BM cells obtained 24 weeks after HSCT from the primary recipients and at 12 weeks after HSCT from the secondary recipients. b Flow cytometry analysis showing the percentage of LSK cells that were of donor CD45.1+/CD45.2+ origin in pooled BM samples from primary recipients (CD45.2+) 24 weeks after transplantation. c Similar analyses conducted in pooled BM samples from secondary recipients (CD45.2+) 12 weeks after transplantation. Bars represent standard error of the mean. *P ≤ 0.05. **P ≤ 0.01. ***P ≤ 0.001. Ad-MSC adipose tissue-derived mesenchymal stem cell, BM bone marrow, HSC hematopoietic stem cell, HSCT hematopoietic stem cell transplant, LSK lineage− Sca-1+ cKit+, ns non-significant difference
Mentions: As observed in previous experiments (Fig. 3), a dose of 6×105 Ad-MSCs significantly increased LSK engraftment in primary recipients during the first weeks after transplant, although at 12 and 24 weeks post-transplantation differences did not reach statistical significance (Fig. 4a). This figure also shows that most of the PB analyses performed in secondary and tertiary recipients revealed improved donor engraftments in the LSK + Ad-MSC group, as compared with the LSK group. Consistent with these observations, the proportion of precursor cells that were of donor origin (percentage of LSK cells that were CD45.1+/CD45.2+) in pooled BM samples that were transplanted in secondary and tertiary recipients was also higher in the LSK + Ad-MSC groups (Fig. 4b, c). These experiments allow us to conclude that the co-infusion of Ad-MSCs improves the engraftment not only of progenitors responsible for the early stages of hematopoietic engraftment but also of long-term repopulating HSCs.Fig. 4

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.