title: A Novel, Single Step, Highly Sensitive In-Vitro Cell-Based Metabolic Assay Using Honeycomb Microporous Polymer Membranes.
abstract: This study describes a novel, simple and versatile system for cell-based assays at the bench-top. The system consists of Polyurethane (PU) based honeycomb membrane with the active compounds/assay reagents dispensed on its pore linings. Membranes with functionalized pores were thus created and used for conducting cell based assays. As proof-of-concept Flourocein acetate (FDA) and Propidium iodide (PI) were embedded on the pore linings and live/dead assays were performed on L929 and Hacat cell lines. The results proved the sensitivity of the membrane based cell assay. To ensure the capacity of this system for high throughput applications, membrane based live/dead assay was performed on L929 cells with varying levels of viability. The results from this experiment were quantified by microscopic and spectrofluourimetric techniques both of which were found to correlate well. It was concluded that this simple membrane based cell assay is highly versatile and enables multiple compounds to be tested on the same cell/tissue. Furthermore, this method requires low volumes of assay reagents and eliminates many of the wet techniques that are involved in a conventional assay, without compromising on the sensitivity. It is anticipated that this functionalized membrane system could be easily adapted for both manual and automated high content screening experiments including in vitro biomaterial evaluation as well as cytotoxicity of nanomaterials.
authors: Shiny
authors: S
authors: Velayudhan
authors: P R Anil
authors: PR
authors: Kumar
authors: Prabha D
authors: PD
authors: Nair
mesh_terms: D000818:Animals; D002460:Cell Line; D045744:Cell Line, Tumor; D002470:Cell Survival; D004305:Dose-Response Relationship, Drug; D015195:Drug Design; D005452:Fluoresceins / Q000737:chemistry; D006801:Humans; D051379:Mice; D049329:Nanostructures / Q000737:chemistry; D036103:Nanotechnology / Q000295:instrumentation* / Q000379:methods*; D011140:Polyurethanes / Q000737:chemistry*; D016062:Porosity; D011419:Propidium / Q000737:chemistry; D013050:Spectrometry, Fluorescence
chemical_list: D005452:Fluoresceins; D011140:Polyurethanes; D011419:Propidium; C018506:diacetylfluorescein