RESUMEN
The complexity of organogenesis hinders in vitro generation of organs derived from a patient's pluripotent stem cells (PSCs), an ultimate goal of regenerative medicine. Mouse wild-type PSCs injected into Pdx1(-/-) (pancreatogenesis-disabled) mouse blastocysts developmentally compensated vacancy of the pancreatic "developmental niche," generating almost entirely PSC-derived pancreas. To examine the potential for xenogenic approaches in blastocyst complementation, we injected mouse or rat PSCs into rat or mouse blastocysts, respectively, generating interspecific chimeras and thus confirming that PSCs can contribute to xenogenic development between mouse and rat. The development of these mouse/rat chimeras was primarily influenced by host blastocyst and/or foster mother, evident by body size and species-specific organogenesis. We further injected rat wild-type PSCs into Pdx1(-/-) mouse blastocysts, generating normally functioning rat pancreas in Pdx1(-/-) mice. These data constitute proof of principle for interspecific blastocyst complementation and for generation in vivo of organs derived from donor PSCs using a xenogenic environment.
Asunto(s)
Blastocisto , Quimera/embriología , Páncreas/citología , Páncreas/embriología , Células Madre Pluripotentes , Animales , Diabetes Mellitus/inducido químicamente , Diabetes Mellitus/terapia , Desarrollo Embrionario , Técnicas de Sustitución del Gen , Proteínas de Homeodominio/genética , Ratones , Ratones Endogámicos , Organogénesis , Ratas , Ratas Wistar , Transactivadores/genéticaRESUMEN
Because a shortage of donor organs has been a major obstacle to the expansion of organ transplantation programs, the generation of transplantable organs is among the ultimate goals of regenerative medicine. However, the complex cellular interactions among and within tissues that are required for organogenesis are difficult to recapitulate in vitro. As an alternative, we used blastocyst complementation to generate pluripotent stem cell (PSC)-derived donor organs in vivo. We hypothesized that if we injected PSCs into blastocysts obtained from mutant mice in which the development of a certain organ was precluded by genetic manipulation, thereby leaving a niche for organ development, the PSC-derived cells would developmentally compensate for the defect and form the missing organ. In our previous work, we showed proof-of-principle findings of pancreas generation by injection of PSCs into pancreas-deficient Pdx1(-/-) mouse blastocysts. In this study, we have extended this technique to kidney generation using Sall1(-/-) mouse blastocysts. As a result, the defective cells were totally replaced, and the kidneys were entirely formed by the injected mouse PSC-derived cells, except for structures not under the influence of Sall1 expression (ie, collecting ducts and microvasculature). These findings indicate that blastocyst complementation can be extended to generate PSC-derived kidneys. This system may therefore provide novel insights into renal organogenesis.
Asunto(s)
Blastocisto/citología , Células Madre Pluripotentes Inducidas/citología , Riñón/crecimiento & desarrollo , Animales , Técnicas de Cultivo de Célula , Diferenciación Celular/fisiología , Células Madre Embrionarias/citología , Genotipo , Riñón/anatomía & histología , Riñón/embriología , Glomérulos Renales/citología , Ratones , Ratones Endogámicos , Ratones Noqueados , Micromanipulación/métodos , Nefronas/citología , Organogénesis/fisiología , Quimera por Trasplante/crecimiento & desarrolloAsunto(s)
Antiinflamatorios/uso terapéutico , Ciclosporina/uso terapéutico , Glomerulonefritis Membranosa/tratamiento farmacológico , Inmunosupresores/uso terapéutico , Prednisolona/uso terapéutico , Relación Dosis-Respuesta a Droga , Quimioterapia Combinada , Humanos , Japón , Estudios Retrospectivos , Resultado del TratamientoAsunto(s)
Nefropatías Diabéticas/complicaciones , Deficiencia del Factor XIII/complicaciones , Fallo Renal Crónico/terapia , Diálisis Renal/métodos , Catéteres de Permanencia , Dalteparina/uso terapéutico , Nefropatías Diabéticas/terapia , Deficiencia del Factor XIII/tratamiento farmacológico , Femenino , Fibrinolisina/uso terapéutico , Humanos , Fallo Renal Crónico/etiología , Persona de Mediana EdadRESUMEN
Graft-versus-host disease (GVHD), mediated by donor-derived alloreactive T cells, is a major cause of nonrelapse mortality in allogeneic hematopoietic stem cell transplantation. Its therapy is not well-defined. We established allele-specific anti-human leukocyte antigen (HLA) monoclonal antibodies (ASHmAbs) that specifically target HLA molecules, with steady death of target-expressing cells. One such ASHmAb, against HLA-A*02:01 (A2-kASHmAb), was examined in a xenogeneic GVHD mouse model. To induce fatal GVHD, non-irradiated NOD/Shi-scid/IL-2Rγ(null) mice were injected with healthy donor human peripheral blood mononuclear cells, some expressing HLA-A*02:01, some not. Administration of A2-kASHmAb promoted the survival of mice injected with HLA-A*02:01-expressing peripheral blood mononuclear cells (p < 0.0001) and, in humanized NOD/Shi-scid/IL-2Rγ(null) mice, immediately cleared HLA-A*02:01-expressing human blood cells from mouse peripheral blood. Human peripheral blood mononuclear cells were again detectable in mouse blood 2 to 4 weeks after A2-kASHmAb administration, suggesting that kASHmAb may be safely administered to GVHD patients without permanently ablating the graft. This approach, different from those in existing GVHD pharmacotherapy, may open a new door for treatment of GVHD in HLA-mismatched allogeneic hematopoietic stem cell transplantation.