Development of a bio-inspired in silico-in vitro platform: towards personalised healthcare through optimisation of a bone-marrow mimicry bioreactor

Human red blood cell production, or erythropoiesis, occurs within bone marrow. Living animal and human cadaver models have demonstrated the marrow production of red blood cells is a spatially-complex process, where cells replicate, mature, and migrate between distinct niches defined by biochemical nutrient access, supportive neighboring cells, and environmental structure. Unfortunately, current research in understanding normal and abnormal human production of blood takes place in petri dishes and t-flasks as 2D liquid suspension cultures, neglecting the role of the marrow environment for blood production. The culture of blood on marrow-mimetic 3D biomaterials has been used as a laboratory model of physiological blood production, but lacks characterization. In this work, a 3D biomaterial platform is developed and to capture the in vivo blood production process and manufacture red blood cells from human umbilical cord blood. First ceramic hollow fibres were designed and tested to be incorporated and perfused in a 3D porous scaffold bioreactor to mimic marrow structure, provide a better expansion of cell numbers, a better diffusion of nutrients, and allow for the continuous, non-invasive harvest of small cells in comparison to static, unperfused biomaterials. Quantitative 3D image analysis tools were developed to spatially assess bioreactor distributions and associations of and between different cell types. Using these tools, the bioreactor distribution of red blood cell production were characterized within niches in collaboration with supportive, non-blood cell types and designed miniaturised, parallelised mini-bioreactors to further explore bioreactor capabilities. This thesis presents a hollow fibre bioreactor able to produce blood cells alongside supportive cells at 1,000-fold higher cell densities with 10-fold fewer supplemented factor than flask cultures, without serum, with one cell source, and continuously harvest enucleate red blood cell product to provide a physiologically-relevant model for cell expansion protocols.Open Acces

Development of a bio-inspired in silico-in vitro platform: Towards personalised healthcare through optimisation of a bone-marrow mimicry bioreactor

Human red blood cell production, or erythropoiesis, occurs within bone marrow. Living animal and human cadaver models have demonstrated the marrow production of red blood cells is a spatially-complex process, where cells replicate, mature, and migrate between distinct niches defined by biochemical nutrient access, supportive neighboring cells, and environmental structure. Unfortunately, current research in understanding normal and abnormal human production of blood takes place in petri dishes and t-flasks as 2D liquid suspension cultures, neglecting the role of the marrow environment for blood production. The culture of blood on marrow-mimetic 3D biomaterials has been used as a laboratory model of physiological blood production, but lacks characterization. In this work, a 3D biomaterial platform is developed and to capture the in vivo blood production process and manufacture red blood cells from human umbilical cord blood. First ceramic hollow fibres were designed and tested to be incorporated and perfused in a 3D porous scaffold bioreactor to mimic marrow structure, provide a better expansion of cell numbers, a better diffusion of nutrients, and allow for the continuous, non-invasive harvest of small cells in comparison to static, unperfused biomaterials. Quantitative 3D image analysis tools were developed to spatially assess bioreactor distributions and associations of and between different cell types. Using these tools, the bioreactor distribution of red blood cell production were characterized within niches in collaboration with supportive, non-blood cell types and designed miniaturised, parallelised mini-bioreactors to further explore bioreactor capabilities. This thesis presents a hollow fibre bioreactor able to produce blood cells alongside supportive cells at 1,000-fold higher cell densities with 10-fold fewer supplemented factor than flask cultures, without serum, with one cell source, and continuously harvest enucleate red blood cell product to provide a physiologically-relevant model for cell expansion protocols