3D bioprinted, vascularized neuroblastoma tumor environment in fluidic chip devices for precision medicine drug testing

Abstract

Neuroblastoma is an extracranial solid tumor which develops in early childhood and still has a poor prognosis. One strategy to increase cure rates is the identification of patient-specific drug responses in tissue models that mimic the interaction between patient cancer cells and tumor environment. We therefore developed a perfused and micro-vascularized tumor-environment model that is directly bioprinted into custom-manufactured fluidic chips. A gelatin-methacrylate/fibrin-based matrix containing multiple cell types mimics the tumor-microenvironment that promotes spontaneous micro-vessel formation by embedded endothelial cells. We demonstrate that both, adipocyte- and iPSC-derived mesenchymal stem cells can guide this process. Bioprinted channels are coated with endothelial cells post printing to form a dense vessel - tissue barrier. The tissue model thereby mimics structure and function of human soft tissue with endothelial cell-coated larger vessels for perfusion and micro-vessel networks within the hydrogel-matrix. Patient-derived neuroblastoma spheroids are added to the matrix during the printing process and grown for more than two weeks. We demonstrate that micro-vessels are attracted by and grow into tumor spheroids and that neuroblastoma cells invade the tumor-environment as soon as the spheroids disrupt. In summary, we describe the first bioprinted, micro-vascularized neuroblastoma – tumor-environment model directly printed into fluidic chips and a novel medium-throughput biofabrication platform suitable for studying tumor angiogenesis and metastasis in precision medicine approaches in future.

Generation of a Perfusable 3D Lung Cancer Model by Digital Light Processing

“…Different strategies have been developed to this end. In many cases, fugitive inks were used that produced a distinct vascular pattern during the printing process and were removed post-printing during the culture period [ 37 , 38 ]. Compared to widely used extrusion-based bioprinting, DLP combines the advantages of extraordinarily high resolution and high stability of the produced constructs.…”

“…In order to improve the physiological relevance of the model further, the channel needs to be seeded with endothelial cells, e.g., HUVECs, in further optimization experiments. Previous studies have shown that a tight endothelial lining can be achieved by post-printing seeding of cells and slow rotation of the model to ensure even distribution of the cells on the entire vessel wall [ 38 ]. The tight endothelium will hinder substances from easily penetrating into the model and thereby more realistically simulate the pharmacokinetic features of human patients.…”

Patient-derived models: Advanced tools for precision medicine in neuroblastoma

“…Functional short-time cultures of both tumor cells and immune cells in a microfluidic system have been reported for adult cancers ( 125 , 126 ) and might be applicable also to pediatric cancers. Very recently, the first bioprinted, vascularized NB microenvironment on a fluidic chip was reported ( 127 ). Implantation of cell line-derived NB spheroids led to NB cell survival for two weeks and successful micro-vessel infiltration of the spheroids ( 127 ).…”

“…Very recently, the first bioprinted, vascularized NB microenvironment on a fluidic chip was reported ( 127 ). Implantation of cell line-derived NB spheroids led to NB cell survival for two weeks and successful micro-vessel infiltration of the spheroids ( 127 ). A different study managed to establish PD NB organotypic slice cultures that could potentially preserve an intact NB tumor microenvironment ( 88 ).…”

3D bioprinting and the revolution in experimental cancer model systems—A review of developing new models and experiences with in vitro 3D bioprinted breast cancer tissue-mimetic structures

“…These models require the reconstitution of the complexity and heterogeneity of glioblastoma and neuroblastoma tissues, and additionally, the special tumour-stroma interactions and blood-brain barriers for accelerating potential therapeutic interventions ( 84 ). These models represent scaffold-free 3D bioprinted spheroid cultures or co-printed, co-cultured glioblastoma or other CNS malignancy-derived tumour cells (e.g., neuroblastoma) cells with the associated macrophages, stromal cells to build the special microenvironment ( 85 , 86 , 87 , 88 ). To develop more reliable models in these diseases, patient-derived newly isolated glioma cells are preferred instead of traditional glioma cell lines which were long cultured in 2D cultures ( 89 ), and another research direction is to combine these with 3D bioprinted blood-brain barrier models ( 90 , 91 ).…”