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Human Alveolar-Like Macrophages Derived from Embryonic Stem Cells for the Treatment and Prevention of Lung Disease
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A7614 - Human Alveolar-Like Macrophages Derived from Embryonic Stem Cells for the Treatment and Prevention of Lung Disease
Author Block: S. Bouch, M. L. Litvack, M. Post; Translational Medicine, The Hospital for Sick Children, Toronto, ON, Canada.
Introduction: Inflammation and dysregulated innate immunity play a vital role in the pathophysiology of many lung diseases. Current clinical practices for treating many of these lung diseases have stagnated in recent years and new immunomodulatory therapeutic approaches are required. To address this our aim was to generate alveolar-like macrophages (ALMs) in vitro from human embryonic stem cells (ESCs). Alveolar macrophages (AMs) are resident cells of the pulmonary innate immune system that are responsible for maintaining healthy airways. Unlike bone marrow-derived macrophages, AMs arise from primitive hematopoiesis and are able to proliferate, making them an ideal candidate for cell-based therapies.
Methods: Using a stage-specific directed differentiation approach we generated human ALMs (hALMs) from H1 ESCs. Under defined media conditions ESCs passed through mesodermal progenitor, hemangioblast, and primitive hematopoiesis stages to produce hALMs. Cells were characterized by flow cytometry for typical myeloid and human AM markers. The functional properties of hALMs were determined by assessing their uptake ability of live P. aeruginosa and acetylated, fluorescently-labeled low-density lipoprotein (DiI-AcLDL). Cryopreservation and recovery capacity was also assessed.
Results: After 1 month of culture hALMs expressed myeloid markers including CD11b, CD11c and CD45, and expressed AM markers CD68, CD163, CD169, CD206 and HLA-DR. These characteristics were maintained after 3 months in culture. hALMs exhibited phagocytic capabilities demonstrated by the uptake of live P. aeruginosa and DiI-AcLDL. In optimum media conditions hALMs exhibited a doubling time of ~48 hours. hALMs undergo cryopreservation and recovery and maintain their cell marker characteristics and proliferative ability.
Conclusion: Together, these data suggest that we are able to successfully generate hALMS derived from ESCs that are capable of phagocytosis and proliferation in vitro, thus proving scalable for future therapeutic applications. Our hALMs are a prime candidate for cell-based therapies to treat lung diseases that result from inflammation and dysregulated innate immunity such as bronchopulmonary dysplasia, cystic fibrosis, chronic obstructive pulmonary disease and infectious lung diseases.
Author Block: S. Bouch, M. L. Litvack, M. Post; Translational Medicine, The Hospital for Sick Children, Toronto, ON, Canada.
Introduction: Inflammation and dysregulated innate immunity play a vital role in the pathophysiology of many lung diseases. Current clinical practices for treating many of these lung diseases have stagnated in recent years and new immunomodulatory therapeutic approaches are required. To address this our aim was to generate alveolar-like macrophages (ALMs) in vitro from human embryonic stem cells (ESCs). Alveolar macrophages (AMs) are resident cells of the pulmonary innate immune system that are responsible for maintaining healthy airways. Unlike bone marrow-derived macrophages, AMs arise from primitive hematopoiesis and are able to proliferate, making them an ideal candidate for cell-based therapies.
Methods: Using a stage-specific directed differentiation approach we generated human ALMs (hALMs) from H1 ESCs. Under defined media conditions ESCs passed through mesodermal progenitor, hemangioblast, and primitive hematopoiesis stages to produce hALMs. Cells were characterized by flow cytometry for typical myeloid and human AM markers. The functional properties of hALMs were determined by assessing their uptake ability of live P. aeruginosa and acetylated, fluorescently-labeled low-density lipoprotein (DiI-AcLDL). Cryopreservation and recovery capacity was also assessed.
Results: After 1 month of culture hALMs expressed myeloid markers including CD11b, CD11c and CD45, and expressed AM markers CD68, CD163, CD169, CD206 and HLA-DR. These characteristics were maintained after 3 months in culture. hALMs exhibited phagocytic capabilities demonstrated by the uptake of live P. aeruginosa and DiI-AcLDL. In optimum media conditions hALMs exhibited a doubling time of ~48 hours. hALMs undergo cryopreservation and recovery and maintain their cell marker characteristics and proliferative ability.
Conclusion: Together, these data suggest that we are able to successfully generate hALMS derived from ESCs that are capable of phagocytosis and proliferation in vitro, thus proving scalable for future therapeutic applications. Our hALMs are a prime candidate for cell-based therapies to treat lung diseases that result from inflammation and dysregulated innate immunity such as bronchopulmonary dysplasia, cystic fibrosis, chronic obstructive pulmonary disease and infectious lung diseases.
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