RESUMEN
PURPOSE: In order to address cell dose limitations associated with the use of cord blood hematopoietic stem cell (HSC) transplantation, we explored the effect of bone marrow stroma-derived heparan sulfate (HS) on the ex vivo expansion of HSCs. METHODS: Heparan sulfate was isolated and purified from the conditioned media of human bone marrow stromal cells and used for the expansion of cord blood-derived CD34(+) cells in the presence of a cocktail of cytokines. RESULTS: The number of myeloid lineage-committed progenitor cells was increased at low dosage of HS as illustrated by an increase in the total number of colony-forming cells (CFC) and colonies of erythroid (BFU-E) and granulocyte-macrophage (CFU-GM) precursors. Notably, the stroma-derived HS did not alter the growth of CD34(+) HSCs or negatively affect the levels of various HSC phenotypic markers after expansion. CONCLUSIONS: This study shows that HS secreted into solution by stromal cells has the capacity to support hematopoietic cytokines in the maintenance and expansion of HSCs. The incorporation of stroma-derived HS as a reagent may improve the efficacy of cord blood HSC transplantation by enhancing the number of committed cells and accelerating the rate of engraftment.
Asunto(s)
Sangre Fetal/citología , Sangre Fetal/efectos de los fármacos , Células Madre Hematopoyéticas/citología , Células Madre Hematopoyéticas/efectos de los fármacos , Heparitina Sulfato/farmacología , Células de la Médula Ósea/química , Recuento de Células , Técnicas de Cultivo de Célula/métodos , Células Cultivadas , Trasplante de Células Madre de Sangre del Cordón Umbilical/métodos , Medios de Cultivo Condicionados , Glicosaminoglicanos/química , Glicosaminoglicanos/aislamiento & purificación , Glicosaminoglicanos/farmacología , Células Progenitoras de Granulocitos y Macrófagos/citología , Células Progenitoras de Granulocitos y Macrófagos/efectos de los fármacos , Heparitina Sulfato/química , Heparitina Sulfato/aislamiento & purificación , Humanos , Células Progenitoras de Megacariocitos y Eritrocitos/citología , Células Progenitoras de Megacariocitos y Eritrocitos/efectos de los fármacos , Células Madre , Células del Estroma/químicaRESUMEN
We report a Jak2V617F knockin mouse myeloproliferative neoplasm (MPN) model resembling human polycythemia vera (PV). The MPN is serially transplantable and we demonstrate that the hematopoietic stem cell (HSC) compartment has the unique capacity for disease initiation but does not have a significant selective competitive advantage over wild-type HSCs. In contrast, myeloid progenitor populations are expanded and skewed toward the erythroid lineage, but cannot transplant the disease. Treatment with a JAK2 kinase inhibitor ameliorated the MPN phenotype, but did not eliminate the disease-initiating population. These findings provide insights into the consequences of JAK2 activation on HSC differentiation and function and have the potential to inform therapeutic approaches to JAK2V617F-positive MPN.
Asunto(s)
Sustitución de Aminoácidos , Células Madre Hematopoyéticas/patología , Janus Quinasa 2/genética , Janus Quinasa 2/metabolismo , Trastornos Mieloproliferativos/genética , Trastornos Mieloproliferativos/patología , Animales , Antígenos CD/metabolismo , Médula Ósea/patología , Células de la Médula Ósea/efectos de los fármacos , Trasplante de Médula Ósea , Recuento de Células , Diferenciación Celular/genética , Modelos Animales de Enfermedad , Células Precursoras Eritroides/metabolismo , Células Precursoras Eritroides/patología , Eritropoyetina/farmacología , Expresión Génica/genética , Perfilación de la Expresión Génica , Hematócrito , Trasplante de Células Madre Hematopoyéticas , Células Madre Hematopoyéticas/efectos de los fármacos , Células Madre Hematopoyéticas/metabolismo , Heterocigoto , Humanos , Janus Quinasa 2/antagonistas & inhibidores , Células Progenitoras de Megacariocitos/metabolismo , Células Progenitoras de Megacariocitos/patología , Células Progenitoras de Megacariocitos y Eritrocitos/efectos de los fármacos , Células Progenitoras de Megacariocitos y Eritrocitos/metabolismo , Células Progenitoras de Megacariocitos y Eritrocitos/patología , Células Progenitoras de Megacariocitos y Eritrocitos/trasplante , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Células Progenitoras Mieloides/metabolismo , Células Progenitoras Mieloides/patología , Trastornos Mieloproliferativos/tratamiento farmacológico , Policitemia Vera/genética , Policitemia Vera/patología , Inhibidores de Proteínas Quinasas/farmacología , Inhibidores de Proteínas Quinasas/uso terapéutico , Pirrolidinas/farmacología , Pirrolidinas/uso terapéutico , Bazo/efectos de los fármacos , Bazo/patología , Sulfonamidas/farmacología , Sulfonamidas/uso terapéutico , Análisis de SupervivenciaRESUMEN
OBJECTIVE: To study the effect of weiganli ([Chinese characters: see text]) on bone marrow hemopoiesis. METHODS: The effects of weiganli on the peripheral blood picture and the number of bone marrow nucleated cells (BMCs) were observed in myelosuppressed anemic model mice, and the effects of weiganli on the growth of colony forming unit-granulocyte macrophage (CFU-GM), colony forming unit-erythroid (CFU-E), burst forming unit-erythroid (BFU-E), colony forming unit-megkaryocyte (CFU-Meg) were investigated by in vitro cell culture technique. The hemopoietic stem cells (HSCs, c-kit+) in bone marrow were double stained with fluorescent antibody PE-C-Kit and FITC-CD45, and the HSCs (c-kit+) were counted by flow cytometer with CD45/SSC (side scatter) gating. RESULTS: Peripheral blood cell counts and the number of BMCs were significantly improved after weiganli administration; and bone marrow hemopoietic stem/progenitor cells were significantly increased. CONCLUSION: Weiganli can effectively promote the recovery of hemopoietic function in the myelosuppressed anemic mice.