Using 3D-bioprinted models to study pediatric neural crest-derived tumors

Abstract

The use of three-dimensional (3D) bioprinting has remained at the forefront of tissue engineering and has recently been employed for generating bioprinted solid tumors to be used as cancer models to test therapeutics. In pediatrics, neural crest-derived tumors are the most common type of extracranial solid tumors. There are only a few tumor-specific therapies that directly target these tumors, and the lack of new therapies remains detrimental to improving the outcomes for these patients. The absence of more efficacious therapies for pediatric solid tumors, in general, may be due to the inability of the currently employed preclinical models to recapitulate the solid tumor phenotype. In this study, we utilized 3D bioprinting to generate neural crest-derived solid tumors. The bioprinted tumors consisted of cells from established cell lines and patient-derived xenograft tumors mixed with a 6% gelatin/1% sodium alginate bioink. The viability and morphology of the bioprints were analyzed via bioluminescence and immunohisto chemistry, respectively. We compared the bioprints to traditional two-dimensional (2D) cell culture under conditions such as hypoxia and therapeutics. We successfully produced viable neural crest-derived tumors that retained the histology and immunostaining characteristics of the original parent tumors. The bioprinted tumors propagated in culture and grew in orthotopic murine models. Furthermore, compared to cells grown in traditional 2D culture, the bioprinted tumors were resistant to hypoxia and chemotherapeutics, suggesting that the bioprints exhibited a phenotype that is consistent with that seen clinically in solid tumors, thus potentially making this model superior to traditional 2D culture for preclinical investigations. Future applications of this technology entail the potential to rapidly print pediatric solid tumors for use in high-throughput drug studies, expediting the identification of novel, individualized therapies.

Applications of 3D Bioprinting Technology to Brain Cells and Brain Tumor Models: Special Emphasis to Glioblastoma

“…116 CaCl 2 was found to be better at maintaining Schwann cell viability compared to BaCl 2 and ZnCl 2 . 116 Many studies on 3D bioprinted GBM 30,31,45,88 and NB models 58,117 that employed alginate-based hydrogels and an extrusion-based strategy utilized CaCl 2 as a cross-linker which has high solubility and induces fast gelation. 112,114 CaSO 4 is reported to make 3D constructs considerably stiffer than other cross-linkers and lead to slow gelation of alginate.…”

“…Furthermore, 3D bioprinted models were found to be more resistant to chemotherapeutic drugs and hypoxia than 2D cell culture rendering 3D bioprinted models more desirable for preclinical trials (Figure 4C,D). 117 While this model is a significant advance for this scope in terms of its superiority to 2D culture and its ability to mimic histology and immunostaining traits of original tumors and continue to grow in mice, the model did not contain noncancerous cells like HUVECS, fibroblasts, or cancer SCs that are highly associated with chemoresistance and disease reoccurrence. 117,141 The 3D bioprinting/printing approach can also be used for other preclinical applications including drug delivery, toxicity efficacy, and assessment of pharmacokinetic parameters.…”

“…48 A very recent study constructed 3D bioprinted neural crestderived solid tumors made of gelatin and sodium alginate with NB cell lines (SK-N-BE(2) and SK-N-AS), NB (COA3 and COA6), and high-grade neuroendocrine-like tumor (COA109) PDXs cells. 117 Two models were created in this study. In the first model, a three-layer model where the top and bottom consisting of hydrogel and the middle layer composed of cells was fabricated (Figure 4A).…”

“…117 While this model is a significant advance for this scope in terms of its superiority to 2D culture and its ability to mimic histology and immunostaining traits of original tumors and continue to grow in mice, the model did not contain noncancerous cells like HUVECS, fibroblasts, or cancer SCs that are highly associated with chemoresistance and disease reoccurrence. 117,141 The 3D bioprinting/printing approach can also be used for other preclinical applications including drug delivery, toxicity efficacy, and assessment of pharmacokinetic parameters. 142 In vitro 3D models have uncovered more biomimetic toxicities in pharmaceuticals than conventional 2D models.…”

“…Many studies on 3D bioprinted GBM ,,, and NB models , that employed alginate-based hydrogels and an extrusion-based strategy utilized CaCl 2 as a cross-linker which has high solubility and induces fast gelation. , CaSO 4 is reported to make 3D constructs considerably stiffer than other cross-linkers and lead to slow gelation of alginate. , CaSO 4 was employed to cross-link electrically conductive nanofibrillated cellulose, alginate, and carbon nanotube-based bioink including SH-SY5Y cells . Electrical conductivity, which was found to enhance neural differentiation, can be adjusted by cross-linking alginate through Ca …”