Biofabrication Of The Brain Glioma Co-culture And Blood Brain Barrier Models In Vitro

Glioma is a malignant tumor that seriously affects human health with strong invasion,infiltrative growth,and vague margin.The deadly consequences of the tumor are not only for its uncontrolled proliferation,but also for complications caused by the interactions with neighboring cells;What`s more,the other problem is the presence of blood-brain barrier,a series of medicines are kept on the outside and unable to achieve the therapeutic effect.Accordingly,this paper proposed the construction of brain glioma co-culture model in vitro by bio-manufacturing method to research the interaction between glioma cells and neighboring cells;On the other hand,a bionic blood brain barrier model was created by the novel composite processes,for the preliminary exploration of drug delivery and cancer treatment.The traditional tumor co-culture model is mostly based on the cell microsphere technology,but this way is difficult to arrange the microstructures such as the relative position of different cells in the microspheres.In order to achieve the better co-culture model with precise and steerable characters,this paper carried out related research based on cell 3D printing technology,it includes the following contents: Firstly,the multi nozzle pneumatic 3D cell printer was design and assembled,with the four section temperature control process,it achieves the high cell survival rate while ensure no salivation,blocking and other undesirable defects.Glioma cells U118 was selected as target cells,brain microvascular endothelial cells and astrocytes as neighboring cells which are two important component in the microenvironment of glioma tumor.Based on the new 3D printer,the co-culture model was formed,and compared with the 2D plane model,3D model that containing one kind cell by the drug test experiment.The result showed that there are significant differences between the three-dimensional and two-dimensional culture environment in morphology and function of cells,and astrocytes promoted the drug stance of glioma cells U118.Secondly,an innovative composite process was adopted to form the bionic blood-brain barrier model,a series of tests and contrast experiments were also carried out,and the results show that: 1),This process has achieved the controlled release of growth factors and induced the endothelial progenitor cells into endothelial cells successfully.Compared with the traditional gold standard Transwell model,the expression of the four specific proteins in this model which are closely related with blood brain barrier`s function have a significant improvement(ALP increased 89.82%,?-GT increased 88.86%,ZO-1 increased 157.4%,Claudin-5 increased 102.32%);2),astrocytes can promote microvascular endothelial cells to form tight junction(ZO-1 increased 249.35%,Claudin-5 increased 184.99%);3),glioma cells(U118)has an obvious destructive effect on the tight junction of blood-brain barrier(ZO-1 decreased 55.86%,Claudin-5 decreased 37.84%).