Neurodegenerative diseases are characterized by the progressive loss of neurons and other brain cells, leading to cognitive or motor disabilities. Brain disorders have a colossal societal and economic burden since about 1 in 3 Europeans are affected by one of these diseases. Available treatments only slow down disease progression or reduce the symptomatology, unfortunately they fail to entirely stop disease progression. The 4-DBR project proposes to develop a 3D bioprinting strategy to obtain a vascularized minibrain, that can be transplanted into patients affected by neurodegenerative disorders. The first step to develop this new strategy is to develop protocols to differentiate the distinct cellular components from the same human Pluripotent Stem Cell (hPSC) line. Thus, we have recently developed a protocol to obtain striatal neurons as one of the main targets of neurodegeneration in Huntington’s disease.

Here we show our recent advances in the production of endothelial cells and pericytes as key components of the blood brain barrier. We show that endothelial cells, expressing both endothelial markers CD31 and von Willebrand factor, can be obtained from hPSC after 10 days of hPSC differentiation. We are currently setting up a MACS purification method. We also show preliminary data of the culture of both endothelial cells and pericytes, growing on decellularized arteries, veins, or brain capillaries as substrate.

Our culture protocols for different cell types serve as a basis in the generation of the bioprinted vascularized minibrain that can be transplanted in patients.