title: Sample preparation strategies for mass spectrometry imaging of 3D cell culture models.
abstract: Three dimensional cell cultures are attractive models for biological research. They combine the flexibility and cost-effectiveness of cell culture with some of the spatial and molecular complexity of tissue. For example, many cell lines form 3D structures given appropriate in vitro conditions. Colon cancer cell lines form 3D cell culture spheroids, in vitro mimics of avascular tumor nodules. While immunohistochemistry and other classical imaging methods are popular for monitoring the distribution of specific analytes, mass spectrometric imaging examines the distribution of classes of molecules in an unbiased fashion. While MALDI mass spectrometric imaging was originally developed to interrogate samples obtained from humans or animal models, this report describes the analysis of in vitro three dimensional cell cultures, including improvements in sample preparation strategies. Herein is described methods for growth, harvesting, sectioning, washing, and analysis of 3D cell cultures via matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) imaging. Using colon carcinoma 3D cell cultures as a model system, this protocol demonstrates the ability to monitor analytes in an unbiased fashion across the 3D cell culture system with MALDI-MSI.
authors.affiliation: Department of Chemistry and Biochemistry, University of Notre Dame; Harper Cancer Research Institute, University of Notre Dame.
authors.forename: Dorothy R
authors.initials: DR
authors.lastname: Ahlf Wheatcraft
authors.affiliation: Department of Chemistry and Biochemistry, University of Notre Dame; Harper Cancer Research Institute, University of Notre Dame.
authors.forename: Xin
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Chemistry and Biochemistry, University of Notre Dame; Harper Cancer Research Institute, University of Notre Dame; ahummon@nd.edu.
authors.forename: Amanda B
authors.initials: AB
authors.lastname: Hummon
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D003110:Colonic Neoplasms / Q000473:pathology; D045325:HCT116 Cells; D006801:Humans; D013048:Specimen Handling / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*