Tumor Cells Develop Defined Cellular Phenotypes After 3D-Bioprinting in Different Bioinks

Abstract

Malignant melanoma is often used as a model tumor for the establishment of novel therapies. It is known that two-dimensional (2D) culture methods are not sufficient to elucidate the various processes during cancer development and progression. Therefore, it is of major interest to establish defined biofabricated three-dimensional (3D) models, which help to decipher complex cellular interactions. To get an impression of their printability and subsequent behavior, we printed fluorescently labeled melanoma cell lines with Matrigel and two different types of commercially available bioinks, without or with modification (RGD (Arginine-Glycine-Aspartate)-sequence/laminin-mixture) for increased cell-matrix communication. In general, we demonstrated the printability of melanoma cells in all tested biomaterials and survival of the printed cells throughout 14 days of cultivation. Melanoma cell lines revealed specific differential behavior in the respective inks. Whereas in Matrigel, the cells were able to spread, proliferate and form dense networks throughout the construct, the cells showed no proliferation at all in alginate-based bioink. In gelatin methacrylate-based bioink, the cells proliferated in clusters. Surprisingly, the modifications of the bioinks with RGD or the laminin blend did not affect the analyzed cellular behavior. Our results underline the importance of precisely adapting extracellular matrices to individual requirements of specific 3D bioprinting applications.

3D Bioprinting of Human Adipose-Derived Stem Cells and Their Tenogenic Differentiation in Clinical-Grade Medium

“…In addition, according to the supplier’s protocols, samples were incubated for characterization and cell culture in fluid media containing calcium to ensure prolonged stability in a watery environment. In accordance with our results, the recently published paper by Schmidt et al qualitatively tested the stability of 3D-printed structures based on a similar alginate and nanocellulose-based bioink, without laminin [ 44 ]. The authors showed that 3D-printed structures starting from this bioink did not show any macroscopically visible changes, suggesting stability of the printed structures up to 14 days and the absence of significant swelling, as demonstrated by the lack of a clear increase in scaffold volume.…”

“…According to our results, Novikova et al reported that MSCs from bone marrow cultured in an alginate hydrogel appeared as atypical cells with spherical shape and inhibited metabolic activity [ 53 ]. Other studies reported that NFC/A hydrogels do not allow or inhibit proliferation and growth of several different types of cells [ 35 , 44 ]. For instance, Müller et al also reported a significant decrease in cell proliferation of chondrocytes embedded in an NFC/A hydrogel [ 35 ].…”

Establishing a 3D In Vitro Hepatic Model Mimicking Physiologically Relevant to In Vivo State

“…A laminin-enriched crosslinkable biomixture provided by Cellink company was used as a bioink. Based on previous studies, laminin-enriched bioink was selected to support the maintenance of hepatic profile expression and clonal expansion of encapsulated cells that have the ability to migrate within bioprinted structures after bioprinting [24,25]. Initially, encapsulated HepG2 cells were well distributed based on the movement of the bioprinter nozzle.…”

Ex Vivo Analysis of Primary Tumor Specimens for Evaluation of Cancer Therapeutics

“…Additional components, such as laminin/peptide additives to Matrigel (Schmidt et al 2019), as well as infiltrating cell types, such as fibroblasts (Amann et al 2014), have been incorporated into semisolid matrices to interact with tumor cells in a way that mimics the in vivo biology of the tumor. These approaches have been used to create ex vivo niches that resemble tumor microenvironments.…”