Friday, July 29, 2016

3DCellMaker for bio-relevant preclinical testing in conventional microplate assays


The PolySciTech product 3DCellMaker (www.3dcellmaker.com) provides a thermogel based platform which allows tumor cells to grow into three-dimensional shapes and structures which have significantly more bio-relevance than 2-dimensional cell monolayers. Multiple studies have shown this to be the case particularly in regards to the micro-environmental difference between the central ‘core’ portion of tumors as compared to the surface environment. This difference is not represented in 2D monolayers and, often, contributes to cancer having resistance to various drugs and chemotherapeutic strategies. Recently, researchers have published regarding protocols for assays of 3D spheroids in microplates by conventional microplate methods. Because 3DCellMaker allows for 3D cell growth in any plasticware or glassware regardless of the container’s dimension, it is well adapted for these types of assays. Read more about these assays here: Vinci, Maria, Sharon Gowan, Frances Boxall, Lisa Patterson, Miriam Zimmermann, Cara Lomas, Marta Mendiola, David Hardisson, and Suzanne A. Eccles. "Advances in establishment and analysis of three-dimensional tumor spheroid-based functional assays for target validation and drug evaluation." BMC biology 10, no. 1 (2012): 1. http://bmcbiol.biomedcentral.com/articles/10.1186/1741-7007-10-29

“Abstract: Background: There is overwhelming evidence that in vitro three-dimensional tumor cell cultures more accurately reflect the complex in vivo microenvironment than simple two-dimensional cell monolayers, not least with respect to gene expression profiles, signaling pathway activity and drug sensitivity. However, most currently available three-dimensional techniques are time consuming and/or lack reproducibility; thus standardized and rapid protocols are urgently needed. Results: To address this requirement, we have developed a versatile toolkit of reproducible three-dimensional tumor spheroid models for dynamic, automated, quantitative imaging and analysis that are compatible with routine high-throughput preclinical studies. Not only do these microplate methods measure three-dimensional tumor growth, but they have also been significantly enhanced to facilitate a range of functional assays exemplifying additional key hallmarks of cancer, namely cell motility and matrix invasion. Moreover, mutual tissue invasion and angiogenesis is accommodated by coculturing tumor spheroids with murine embryoid bodies within which angiogenic differentiation occurs. Highly malignant human tumor cells were selected to exemplify therapeutic effects of three specific molecularly-targeted agents: PI-103 (phosphatidylinositol-3-kinase (PI3K)-mammalian target of rapamycin (mTOR) inhibitor), 17-N-allylamino-17-demethoxygeldanamycin (17-AAG) (heat shock protein 90 (HSP90) inhibitor) and CCT130234 (in-house phospholipase C (PLC)γ inhibitor). Fully automated analysis using a Celigo cytometer was validated for tumor spheroid growth and invasion against standard image analysis techniques, with excellent reproducibility and significantly increased throughput. In addition, we discovered key differential sensitivities to targeted agents between two-dimensional and three-dimensional cultures, and also demonstrated enhanced potency of some agents against cell migration/invasion compared with proliferation, suggesting their preferential utility in metastatic disease. Conclusions: We have established and validated a suite of highly reproducible tumor microplate three-dimensional functional assays to enhance the biological relevance of early preclinical cancer studies. We believe these assays will increase the translational predictive value of in vitro drug evaluation studies and reduce the need for in vivo studies by more effective triaging of compounds. Keywords: 3D angiogenesis drug response high throughput invasion migration tumor spheroids”
Post a Comment