A modified 384‐well‐device for versatile use in 3D cancer cell (co‐)cultivation and screening for investigations of tumor biology in vitro

Abstract

Pancreatic cancer exhibits a worst prognosis owed to an aggressive tumor progression i.a. driven by chemoresistance or tumor‐stroma‐interactions. The identification of candidate genes, which promote this progression, can lead to new therapeutic targets and might improve patient's outcome. The identification of these candidates in a plethora of genes requires suitable screening protocols. The aim of the present study was to establish a universally usable device which ensures versatile cultivation, screening and handling protocols of cancer cells with the 3D spheroid model, an approved model to study tumor biology. By surface modification and alternative handling of a commercial 384‐well plate, a modified device enabling (i) 3D cultivation either by liquid overlay or by a modified hanging drop method for (ii) screening of substances as well as for tumor‐stroma‐interactions (iii) either with manual or automated handling was established. The here presented preliminary results of cell line dependent dose‐response‐relations and a stromal‐induced spheroid‐formation of the pancreatic cancer cells demonstrate the proof‐of‐principle of the versatile functionality of this device. By adapting the protocols to automation, a higher reproducibility and the ability for high‐throughput analyses were ensured.

Spheroids and organoids as humanized 3D scaffold‐free engineered tissues for SARS‐CoV‐2 viral infection and drug screening

“…Spheroids can be produced by different 3D cell culture techniques 35,40 . Different studies have already been published showing high‐throughput production of spheroids from different cell types by using the hanging‐drop technique 61‐65 . Other techniques already describe high‐throughput spheroid production consisting of using nonadhesive agarose microwell systems, 66 cell culture plates with a higher number of wells, as described by Grinner et al, 67 which used a 1536‐well plate format to engineer tumor spheroids for drug screening tests.…”

“…35,40 Different studies have already been published showing high-throughput production of spheroids from different cell types by using the hanging-drop technique. [61][62][63][64][65] Other techniques already describe high-throughput spheroid production consisting of using nonadhesive agarose microwell systems, 66 cell culture plates with a higher number of wells, as described by Grinner et al, 67 which used a 1536-well plate format to engineer tumor spheroids for drug screening tests. Other studies were already performed using well plates for high-throughput applications, as performed by Liao and collaborators 68 ; however, these authors used a 96well plate to cast agarose microwells.…”

Devices and techniques used to obtain and analyze three‐dimensional cell cultures

“…In roundbottomed well plates coated with either agar or poly-2-hydroxyethyl methacrylate (poly-HEMA) (both methods share similar limitations), cell suspensions are seeded, and the plates are subsequently centrifuged to encourage co-localization and adhesion of cells. 4,26 Some cell lines (MDA-MB-231, MDA-MB-468, SKBR3, MDA-MB-361) form loose aggregates when this technique is used; addition of liquid reconstituted basement membrane has circumvented this problem. 11 Another limitation of the forced floating technique is the time required to coat the plates prior to cell seeding, autoclaving of agar before application and preparation of poly-HEMA solution, which can take a few hours.…”

“…Cultures from 96-well plates, 384-well plates and higher have can be used for high-throughput testing of drug efficacy and toxicity studies, gene expression profiling and several biochemical analyses. 26,27…”

The 3D Cell Culture System in the Study of Tumor-Applications and Prospects

“…Most scholars are currently working on establishing a universal 3D cancer culture and screening device to ensure a high degree of flexibility in selecting well-developed 3D culture methods and screening methods for any cell sample. Previous experiments by Miriam Widder, a modified 384-well-device for investigations of tumor biology was established, through higher reproducibility and the ability for high-throughput analyses, the screening and treatment of cancer cells have a good effect [15]. Now, the most popularly used 3D tumor model is a multicellular sphere.…”