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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.


Influence of Ad-MSC dose fractionation on the engraftment of purified HSCs. a Illustration of the experimental protocol corresponding to results shown in b. As a control group, 5 Gy-irradiated mice were intravenously infused with 1500 LSK cells on day 0. Three study groups received one to three different Ad-MSC doses intravenously co-infused, following different administration schemes, depicted in the figure’s experimental design. b Analysis of the donor cell engraftment (fold-increase) with respect to mice transplanted with 1500 LSK cells. Bars represent standard error of the mean. *P ≤ 0.05. ***P ≤ 0.001. Ad-MSC adipose tissue-derived mesenchymal stem cell, HSC hematopoietic stem cell, LSK lineage− Sca-1+ cKit+, ns non-significant difference
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Fig6: Influence of Ad-MSC dose fractionation on the engraftment of purified HSCs. a Illustration of the experimental protocol corresponding to results shown in b. As a control group, 5 Gy-irradiated mice were intravenously infused with 1500 LSK cells on day 0. Three study groups received one to three different Ad-MSC doses intravenously co-infused, following different administration schemes, depicted in the figure’s experimental design. b Analysis of the donor cell engraftment (fold-increase) with respect to mice transplanted with 1500 LSK cells. Bars represent standard error of the mean. *P ≤ 0.05. ***P ≤ 0.001. Ad-MSC adipose tissue-derived mesenchymal stem cell, HSC hematopoietic stem cell, LSK lineage− Sca-1+ cKit+, ns non-significant difference

Mentions: To investigate whether the HSC engraftment effects mediated by Ad-MSCs require their co-infusion with the LSK cells, two different sets of experiments were conducted. In the first experiments (Fig. 6), a group of recipient mice was co-infused with 1500 purified LSK cells together with Ad-MSCs, as in our previous experiments. Additional groups of transplanted mice were now included, in which the total Ad-MSC dose was split into up to three infusions that were administered either the day before or both the day before and the day after the co-infusion of LSK and Ad-MSCs (see experimental protocol in Fig. 6a). As shown in Fig. 6b, doses of 6×105 and 106 Ad-MSCs—which markedly improved the engraftment of 1500 LSK cells when co-infused in a single dose—had no effect when divided into three administrations, and only one of them co-infused with the LSK cells. These studies thus indicate that the total effective dose of Ad-MSCs need to be infused with the LSK cells at the moment of the LSK infusion but not earlier or later than LSK infusion.Fig. 6


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)

Influence of Ad-MSC dose fractionation on the engraftment of purified HSCs. a Illustration of the experimental protocol corresponding to results shown in b. As a control group, 5 Gy-irradiated mice were intravenously infused with 1500 LSK cells on day 0. Three study groups received one to three different Ad-MSC doses intravenously co-infused, following different administration schemes, depicted in the figure’s experimental design. b Analysis of the donor cell engraftment (fold-increase) with respect to mice transplanted with 1500 LSK cells. Bars represent standard error of the mean. *P ≤ 0.05. ***P ≤ 0.001. Ad-MSC adipose tissue-derived mesenchymal stem cell, HSC hematopoietic stem cell, LSK lineage− Sca-1+ cKit+, ns non-significant difference
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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

Fig6: Influence of Ad-MSC dose fractionation on the engraftment of purified HSCs. a Illustration of the experimental protocol corresponding to results shown in b. As a control group, 5 Gy-irradiated mice were intravenously infused with 1500 LSK cells on day 0. Three study groups received one to three different Ad-MSC doses intravenously co-infused, following different administration schemes, depicted in the figure’s experimental design. b Analysis of the donor cell engraftment (fold-increase) with respect to mice transplanted with 1500 LSK cells. Bars represent standard error of the mean. *P ≤ 0.05. ***P ≤ 0.001. Ad-MSC adipose tissue-derived mesenchymal stem cell, HSC hematopoietic stem cell, LSK lineage− Sca-1+ cKit+, ns non-significant difference
Mentions: To investigate whether the HSC engraftment effects mediated by Ad-MSCs require their co-infusion with the LSK cells, two different sets of experiments were conducted. In the first experiments (Fig. 6), a group of recipient mice was co-infused with 1500 purified LSK cells together with Ad-MSCs, as in our previous experiments. Additional groups of transplanted mice were now included, in which the total Ad-MSC dose was split into up to three infusions that were administered either the day before or both the day before and the day after the co-infusion of LSK and Ad-MSCs (see experimental protocol in Fig. 6a). As shown in Fig. 6b, doses of 6×105 and 106 Ad-MSCs—which markedly improved the engraftment of 1500 LSK cells when co-infused in a single dose—had no effect when divided into three administrations, and only one of them co-infused with the LSK cells. These studies thus indicate that the total effective dose of Ad-MSCs need to be infused with the LSK cells at the moment of the LSK infusion but not earlier or later than LSK infusion.Fig. 6

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.