Notes: To demonstrate the differences between 2D cultures and 3D cultures of various complexities, models were set up for three pathologies: breast (MCF7), prostate (LNCaP) and lung (H1437) carcinomas. The models consisted of mono- or co-cultures of established cell lines with fibroblast cell lines. Co-culture fibroblasts were tissue-derived cancer-associated cell lines with the exception of breast cancer where no CAF was available. MCF7 cells were co-cultured with human dermal fibroblasts. Models were microspheroids free floating in plate wells, alginate microspheroids in bioreactors, and cells on an extracellular matrix. All cell lines were stably transfected with a fluorescent protein and used fluorescence to monitor cell proliferation. CellTiter-Glo® 3D Cell Viability Assay was used to viability of the mono- and co-cultures of alginate microspheres grown in the bioreactors. The method employed for use was quite intensive: 40 minutes under agitation, pipetting 10 times, followed by 40 more minutes of agitation before reading the luminescence.  (4818)

Notes: The CellTiter-Glo® 3D Cell Viability Assay was used to measure viability of MDA-MB-468 cells grown on Matrigel with inducible knockdown of PNUTS. The effect of PNUTS knockdown was also assessed in a reporter model of E-Cadherin expression using a firefly luciferase reporter vector and Renilla luciferase control vector. Luciferase activities were measured with the Dual-Luciferase® Reporter Assay System. (4823)

Notes: The authors were investigating new methods of biofabrication of 3D multicellular constructs.  Cells were incorporated into a macroporous scaffold, encapsulated with alginate and printed onto a cold surface with agarose.  The viability immediately after printing was monitored hourly over 72 hours with the RealTime-Glo™ MT Cell Viability Assay. Total cell viability in the 3D constructs was monitored at day 2, 7 and 14 with the CellTiter-Glo® 3D Cell Viability Assay. (4819)

Notes: Several methods of making cryogels were evaluated for the ability of LNCaP cells to colonize and proliferate. LNCaP cell viability was monitored with the CellTiter-Glo® 3D Cell Viability Assay.  (4812)

Notes: SH-SY5Y cells were either plated above or incorporated into hydrogels containing an amyloid protein or collagen. The 3D cultures (incorporated into the hydrogels) viability was measured with the CellTiter-Glo® 3D Cell Viability Assay.  (4820)

Notes: Several matrices (Matrigel, Cultrex Basement Membrane Extract and Alvetex plates) were evaluated for growth of DU145 and LNCaP cells in comparison to 2D culture. For DU145 cultures, Alvetex plates and 2D cultures were very similar in cell viability at 72 hours but 2D overwhelmed the 3D methods by 120 hours. The DU145 cells barely grew on the Matrigel or Basement Membrane Extract.   For LNCaP cells, all were similar at 72 hours but 2D was best at 120 hours with Matrigel and Alvetex plates at 50–60%.The different culture methods were also evaluated for compound toxicity. 2D cultures of DU145 were most susceptible to Docetaxel but mostly insensitive to rapamycin. Alvetex plates grown DU145 were insenstive to rapamycin but Matrigel and basement membrane extract showed some sensitivity.  LNCaP cells grown in all four conditions were susceptible to docataxel and rapamycin with 2D and Alvetex-grown cells being most susceptible. Cell viability in all four growth conditions was measured with the CellTiter-Glo® 3D Cell Viability Assay. (4813)

Notes: The CellTiter-Glo® 3D Cell Viability Assay was used to monitor viable MDA-MB-231 or MMTV-myc cells grown on Matrigel in 96-well plates. (4822)

Notes: The effect of miR-124 on the Nur77-3’ UTR was analyzed with a dual-reporter plasmid and luciferase activities were measured with the Dual-Glo® Luciferase Assay System. Daoy cells used in the study were transfected with the reporter plasmid and miRNA expression vectors using the FuGENE® 6 Transfection Reagent. FuGENE® 6 was also employed to transfect 293T cells for recombinant lentivirus construction. RNA isolation of mRNA expression level determination by probe-based RT-qPCR was accomplished with a Maxwell® 16 Research Instrument using the Maxwell® 16 LEV simplyRNA Tissue Kit. Viability assays of 2D cultures of Daoy cells were determined with the CellTiter-Glo® Cell Viability Assay. Viability assays of 3D microspheroid cultures of Daoy cells (presumably ULA plate-derived) were measured with the CellTiter-Glo® 3D Cell Viability Assay. (4816)

Notes: The authors of this study sought to determine whether cucurbitacin B has antitumor activity against the ovarian cancer cell line A2870 and whether it can sensitize the cisplatin-resistant cell line A2780CP to treatment. They compared caspase activity in A2780CP cells either preincubated with cucurbitacin B before treatment with cisplatin or cells not pretreated using the Caspase-Glo® 3/7 Caspase Assay. Oxidized and total glutathione were measured from both cell lines (before and after cisplatin exposure, with and without preincubation with cucurbitacin B) using the GSH/GSSH-Glo™ Glo Assay. The level of reactive oxygen species in cell lines was also measured by detecting ROS converted to H₂O₂ using the ROS-Glo™ Assay. Cell Viabilty of 3D spheroids formed from the A2780CP cell line was assessed using the CellTiter-Glo® 3D Cell Viability Assay. (4581)

Notes: The authors of this study use large 3D multipcellular spheroids (MCSs, >200µm) to serve as intermediate models between 2D cell culture and in vitro systems for investigating drug delivery and photodynamic therapy (PDT) for cancer. They evaluate four metal complexes that target the mitocondrion for PDT in 3D MCSc formed from HeLa cells. Cell viability is evaluated in the standard 2D culture system using the CellTiter-Glo® Cell Viability Assay, and in the 3D MCS using the CellTiter-Glo® 3D Cell Viability Assay. (4583)

Notes: The authors used a 3D cell culture model to investigate the role of collagen destruction in the pathogenesis of tuberculosis. The CellTiter-Glo® 3D Cell Viability Assay was used to assess cell viability in microspheres formed from peripheral blood mononuclear cells that had been infected with UV-killed M. tuberculosis, and the assay results were read on the GloMax® Discover instrument. (4582)

Notes: The authors used the CellTiter-Glo™ 3D Cell Viability Assay to measure viability of microspheres containing PBMCs infected with UV-killed M. tuberculosis. Luminescence was measured on the GloMax® Discover. (4764)

Notes: The authors of this study used the CellTiter-Glo® 3D Cell Viability Assay to assess the ATP content of HCT-116-derived microtissue spheroids after a bioactivation experiment performed in a microfluidic hanging drop network. For this experiment, the authors designed an array containing three columns of hanging drop microtissue spheroids derived from freshly isolated rat liver cells and a fourth column containing hanging drop microtissue spheroids derived from colorectal carcinoma cells (HCT-116 expressing GFP). The array was perfused with a anticancer prodrug, cyclophosphamide, which gains efficacy after bioactivation by cytochrome P450 enzymes in liver. The authors were able to demonstrate bioactivation of the drug and measure reduced cell viability in the HCT-116-derived microtissue spheroids. (4580)