Three-Dimensional Cell Culture Based on Magnetic Fields to Assemble Low-Grade Ovarian Carcinoma Cell Aggregates Containing Lymphocytes

Abstract

There is a limited number of established ovarian cancer cell lines matching the low-grade serous histotype available for research purposes. Three-dimensional (3D) culture systems provide in vitro models with better tissue-like characteristics than two-dimensional (2D) systems. The goal in the study was to characterize the growth of a given low-grade serous ovarian carcinoma cell line in a 3D culture system conducted in a magnetic field. Moreover, the culture system was evaluated in respect to the assembly of malignant cell aggregates containing lymphocytes. CAISMOV24 cell line alone or mixed with human peripheral blood mononuclear cells (PBMC) were cultured using a commercially available 3D culture system designed for 24 well plates. Resulting cell aggregates revealed the intrinsic capacity of CAISMOV24 cells to assemble structures morphologically defined as papillary, and reflected molecular characteristics usually found in ovarian carcinomas. The contents of lymphocytes into co-cultured cell aggregates were significantly higher (p < 0.05) when NanoShuttle-conjugated PBMC were employed compared with non-conjugated PBMC. Moreover, lymphocyte subsets NK, T-CD4, T-CD8 and T-regulatory were successfully retrieved from co-cultured cell aggregates at 72h. Thus, the culture system allowed CAISMOV24 cell line to develop papillary-like cell aggregates containing lymphocytes.

Magnetic-Based Human Tissue 3D Cell Culture: A Systematic Review

“…Magnetic-based models allow us to effectively produce homogeneous 3D aggregates and with desirable compactness [ 1 , 17 ]. Of the twenty-five studies included in this review, nine studies used only magnetic levitation [ 17 , 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 ], eleven studies used only the bioprinting method [ 14 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 ], and three studies used both methods [ 15 , 36 , 37 ]. Only two studies used the magnetic ring formation method [ 22 , 38 ].…”

“…The 24-well plates were the most used for studies using the levitation method; whereas, the 96-well plates were the most used for the bioprinting method. More precisely, nine studies used the 24-well Bio-Assembler Kit [ 15 , 23 , 24 , 25 , 26 , 30 , 32 , 37 , 39 ], eight studies used the 96-well Bioprinting Kit [ 14 , 27 , 28 , 29 , 31 , 33 , 35 , 36 ], two studies used both 6-well and 24-well Bio-Assembler Kit [ 17 , 19 ], one study used only the 6-well Bio-Assembler Kit [ 34 ], and one study used the 96-well Ring Drive for ring structure formation [ 38 ]. There is an additional study that used tissue culture Petri dishes covered by a top cover with an attached neodymium magnet [ 22 ].…”

“…The use of mouse preadipocyte cells and murine endothelial cells allowed for the formation of adipospheres as a model of white adipose tissue development and growth [ 19 ]. Heterotypic m3D cultures constituted by human ovarian cancer cells (CAISMOV24 cell line) and peripheral blood mononuclear cells (PBMCs), such as monocytes, lymphocytes, and macrophages, were used to study the growth of a low-grade serous ovarian carcinoma and the role of the interactions with immune cells in the tumor microenvironment [ 15 ]. Pancreatic tumor cells isolated from KPC-transgenic mice and murine embryonic fibroblasts were combined in m3D cultures to study ultrasound therapy [ 30 ].…”

“…These advantages have driven the development of studies using m3D-based cultures over the years for a variety of research purposes [ 15 ]. Therefore, this systematic review aims to bring together studies focused on m3D-based cell cultures to explore the different 3D models already developed with this technology, including tumor models, as well as the characteristics of the 3D structures formed, the most suitable materials to use, the implemented analysis techniques, and the limitations and possible applicability of these methods to create heterotypic 3D cultures.…”

Case Report: Formation of 3D Osteoblast Spheroid Under Magnetic Levitation for Bone Tissue Engineering

“…The charged cells can then levitate over any stiff substrate by using neodymium magnetic excitators; the cells aggregate towards the air-liquid interface and work with each other to build a larger 3D structure that involves protein synthesis such as collagen, fibronectin, and laminin ( Tseng et al, 2013 ). This magnetic levitation system has been reported as non-toxic and does not affect proliferation or induce any inflammatory response by cultured cells and is a well-established system culture onto tumor cell lines and primary cells as endothelial, smooth muscle, hepatocytes, and fibroblast ( Souza et al, 2010 ; Tseng et al, 2014 ; Tseng et al, 2018 ; Natânia de Souza-Araújo et al, 2020 ; Urbanczyk et al, 2020 ). Thus, the reported technique is an alternative to both scaffold-based and scaffold-free 3D cell culture systems.…”