RESUMO
Surge protector: a two-component peroxynitrite-generating platform has been engineered to release peroxynitrite (PN) in situ under the control of light. The system, which is constructed by layering sol-gel matrices containing xanthine oxidase (bottom layer) and a metal nitrosyl (top layer), allows studies of PN chemistry at varying fluxes of its precursors.
Assuntos
Luz , Ácido Peroxinitroso/síntese química , Materiais Biomiméticos/química , Materiais Biomiméticos/metabolismo , Géis/química , Concentração de Íons de Hidrogênio , Manganês/química , Óxido Nítrico/química , Oxirredução , Ácido Peroxinitroso/química , Espectrometria de Fluorescência , Superóxidos/química , Raios Ultravioleta , Xantina Oxidase/química , Xantina Oxidase/metabolismoRESUMO
Cell-based assays performed in multiwell plates are utilized in basic and translational research in a variety of cell models. The assembly of these multiwell platforms and their use is often laboratory specific, preventing the standardization of methods and the comparison of outputs across different analytical sites. Moreover, when cell models are based on primary cells with specialized culture requirements, including three-dimensional (3D) cell culture, their complexity and the need for manipulation by experienced operators can add significant cost and introduce long lead times to analysis, both of which are undesirable in any preclinical situation. To address this issue, we explored adaptations of cryopreservation technology that allow cells to be cryopreserved in-plate, ready for use in analysis, and have developed a method applicable to cells from different origins and different culture formats. Here we describe the application of this technology to conventional two-dimensional (2D) monolayers of human mesenchymal stem cells (MSCs) and human macrophages derived from primary monocytes, and to 3D cultures of hepatic organoids, colon organoids, and colon tumor organoids, each presented for cryopreservation in their obligate extracellular matrix. We demonstrated that cell viability, cell physiology, and cytotoxic sensitivity were maintained after cryopreservation, such that the models offer the means to uncouple model assembly from analytical use and to standardize cell models in product form for distribution to end users.