Notes: These authors evaluated the impact of spheroid age on drug efficacy screening to better understand variability from lab to lab when using similar models. HepG2 spheroids were established and evaluated for parameters including viability and CYP3A4 activity over an 18-day period. (5210)

Notes: Multicellular human iPSC liver organiods were generated and assessed using methods including cell viability and CYP3A4 activity assays. (5208)

Notes: This article describes a novel 3D tissue model mimicking pancreatic cancer tumor microenvironment using three different cell types. Cell viability in response to therapeutics was analyzed using the CellTiter-Glo® 3D Cell Viability Assay. The authors present a novel approach for testing therapeutic efficacy in pancreatic cancer. (5182)

Notes: Human pancreatic epithelial carcinoma cells were dispensed at 2,500 cells/well with 20µl of medium in 384-well 3D spheroid culture plates, the plates centrifuged and incubated for 1 day. After verifying spheroid formation, test or control compounds were added the cultured cells, incubated for 24 hours and 20µl of CellTiter-Glo® 3D Cell Viability Assay Reagent was added. Luminescence was measured after 30 minutes. BxPC-3-KRASG12V and BxPC-3-KRASWT cells were seeded into 384-well plates at a density of 2,500 cells/well in 20µl of medium as 3D or 2D cultures. After 24 hours, test compounds or vehicle are added followed by the RealTime-Glo™ Annexin V Apoptosis and Necrosis Assay Reagent and luminescence monitored for 24 hours. (4971)

Notes: These authors engineered three-dimensional cultures of human fibroatheroma (ps-plaque) using a hanging-drop method. They used flow cytometry analysis to study the myeloid cell populations in ps-plaques compared to those in plaques isolated from human carotid arteries. The CellTiter-Glo® 3D Assay was used to assess cell viability within the 3D plaque cultures. (5023)

Notes: These authors used a 3D printer to create cardiac patches using human induced pluripotent stem cell-derived cardiomyocytes, fibroblasts and endothelial cells aggregated to create mixed cell spheroids. Cell viability in the spheroids was determined using the CellTiter-Glo® 3D Assay. The DeadEnd™ Fluorometric TUNEL System was used to assess viability in cut sections of the 3D bioprinted cardiac tissue.  (5024)

Notes: These authors suggest that claudin-2, which is highly expressed in human lung adenocarcinoma cells and is involved in the up-regulation of cell proliferation, may be a novel therapeutic target in lung adenocarcinoma. Claudin-2 knockdown increased the accumulation of anticancer agents in cancer cells and spheroids. As part of the study, the CellTiter-Glo® 3D Assay was used to assess viability of Claudin-2KD/A549 spheroid cultures. (5021)

Notes: FTC236 and C643 thyroid cancer cells were grown as microspheroids in ULA plates. Spheroid growth was promoted by shRNA knockdown of RCAN1-4 in contrast to 2D cultures, which showed no effect. Overexpression of RCAN1-4 lead to greater sensitivity to doxycycline treatment. 3D Cell growth was monitored with the CellTiter-Glo® 3D Cell Viability Assay. (4821)

Notes: The CellTiter-Glo® 3D Cell Viability Assay was used to determine the IC₅₀ values. (4883)

Notes: Cell viability was assessed using the CellTiter-Glo® 3D assay. (4884)

Notes: Human PBMC were isolated and encapsulated in a alginate-collagen matrix (via electrostatic bead generator) either alone or after infection with Mycobacterium tuberculosis. Viability of PBMC-alone or M. tuberculosis-infected PBMC microspheres were measured with the CellTiter-Glo® 3D Cell Viability Assay. Luminescence measurements in the study were accomplished with either the GloMax® 20/20 Luminometer or GloMax® Discover Instrument. (4815)

Notes: ARPE-19 cells were seeded at 1 × 10⁴ cells per well in 96-well plates and incubated for 12 hours. Cells were treated with  H₂O₂ with or without olparib for 4 hours. Cellular NAD+ levels were measured with the NAD/NADH-Glo™ Assay.  Cellular ATP levels were measured with the CellTiter-Glo® Assay. (28292900)

Notes: These authors present a method for generating large and homogenous cell spheroids using Huh7 hepatocellular carcinoma cells. They used the CellTiter-Glo® 3D Assay to determine viability of the cells within the spheroids. (5025)

Notes: Human amniotic fluid mesenchymal stem cells were seeded in a human-derived collagen hydrogel and in a bovine-derived collagen sheet at 3 matching densities. Cell viability was analyzed at 1, 3, and 5 days using the CellTiter-Glo® 3D Cell Viability Assay. (4824)

Notes: This study investigated the effect of the hepatotoxicant and hepatocarcinogen methapyrilene hydrochloride on iron metabolism in rat liver in an in vivo toxicity study and in human HepaRG cell cultures. Cell viability was assessed using the CellTiter-Glo® 3D Assay. The resulting dose-response curve was used to determine the IC10 and IC25 values. (5026)

Notes: These authors sought to develop a three-dimensional culture system that mimicked the embryonic palate and could be used to study fusion behavior in vitro. They used the CellTiter-Glo® 3D Assay to determine cellular ATP content in the 3D spheroid cultures. (5027)

Notes: Primary pancreatic cancer cell lines were grown a microsphere by culturing in round bottom plates in the presence of methylcellulose. Spheroids were assessed for sensitivity to JQ1 using the CellTiter-Glo® 3D Cell Viability Assay. Cell lines expressing high levels of c-myc were sensitive to JQ1. RNA was isolated from cell lines and targeted expression analysis was performed by reverse transcription with the GoScript™ Reverse Transcription System followed by dye-based qPCR with the GoTaq® qPCR Master Mix on the AriaMx real-time PCR instrument (Agilent). (1825)

Notes: Adenoma-derived organoids were established in Matrigel in 96--well plates at 10,000 cells per well. Cells were treated with a γ-secretase inhibitor or vehicle.  Growth of organoids was tremendously reduced compared to vehicle control after 5 days of treatment.  Cell viability was measured with the CellTiter-Glo® 3D Cell Viability Assay. Luminescence was measured with a GloMax® Instrument. (4814)

Notes: Optimal cryogel composition was determined and seeded with 500 LNCaP cells and studied over 3 weeks in culture. The viability was monitored at day 0, 7, 14 and 21 with the CellTiter-Glo® 3D Cell Viability Assay. (4817)

Notes: To quantify 3D spheroids viability, researchers in this study measured ATP content using the CellTiter-Glo® 3D Cell Viability Assay. (4882)

Notes: The CellTiter-Glo® 3D Cell Viability Assay, Perfecta3D Assay (3Dbiomatrix, Inc.) and simple Trypan Blue exclusion were assessed for measuring cytotoxicity of A549 cell microspheroids. The authors state, “The best and most reproducible method to determine the viability of large spheroids for both chemical and for physical treatments was the CellTiter-Glo 3D Cell Viability assay…” The CellTiter-Glo® 3D Assay reported a linear relationship for spheroids that were 350–650 microns with some deviation from linear for spheroids with a diameter of 850 microns. The authors note that this could be related to physical interference with such large spheroids as well as the presence of a necrotic core in the large spheroids. The luminescence from the CellTiter-Glo® 3D Assays was measured with a GloMax® Instrument. (4642)

Notes: The glioblastoma cell lines U-251 MG (containing mutant p53) and U-87 MG (containing wildtype p53) were grown as microspheroids on ultra-low attachment 96-well plates and treated with monocarboxylate transporter (MCT) 1 inhibitors. Morphology was examined, and viability determined by transferring well contents to white 96-well plates and assaying with the CellTiter-Glo® 3D Cell Viability Assay. Luminescence was measured using a GloMax® Discover Instrument. Relative viability was compared to the same cells grown under monolayer conditions. Cells grown under 3D conditions were more susceptible to the MCT inhibitors. (4643)

Notes: Normal human dermal fibroblasts were allowed to form microspheres. Eight microspheroids were seeded per well of a 24-well plate containing electrospun polyurethane. Microspheroids were also placed into wells of a standard tissue-culture-treated plate to serve as a positive control for 100% viability. Cells were cultured for 24 or 72 hours, incubated with CellTiter-Glo® 3D Reagent, and supernatants transferred to a 96-well plate for detection in a 96-well luminometer. Microspheroids cultured on the polyurethane were judged to be 95% viable. (4641)

Notes: Topoisomerase IIα was identified as an important factor for metastasis of colorectal cancer involving the wnt signaling pathway. Microspheroids of SW620 cells, generated through ultra-low attachment plates, were used in the study. Activation of the wnt pathway was monitored with a reporter assay measured with the ONE-Glo™ Luciferase Assay System. Duplicate cells were monitored for viability with the CellTiter-Glo® 3D Cell Viability Assay. An ATP-competitive, N-terminal inhibitor of topoisomerase IIα was examined for cytotoxicity to the microspheroids. Cytotoxicity was monitored from 6–72 hours of exposure to the inhibitor using the CellTox™ Green Cytotoxicity Assay. Cytotoxicity increased over time with the inhibitor, and controls were essentially unaffected. (4708)

Notes: CellTiter-Glo® Cell Viability Assay was used to assess the vulnerability of myc-driven cancers to inhibitors of the NAMPT. Sensitivity was correlated to the level of myc expression. Apoptosis was assessed with the Caspase-Glo® 3/7 Assay. The NAD/NADH-Glo® Assay was used to provide a quantitative measure of NAD+ in the cells under study. (4835)