Imaging Mass Spectrometry of Three-Dimensional Cell Culture Systems

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• 作者：Haohang Li ; Amanda B. Hummon
• 刊名：Analytical Chemistry
• 出版年：2011
• 出版时间：November 15, 2011
• 年：2011
• 卷：83
• 期：22
• 页码：8794-8801
• 全文大小：981K
• 年卷期：v.83,no.22(November 15, 2011)
• ISSN：1520-6882

文摘

Three-dimensional (3D) cell cultures have increased complexity compared to simple monolayer and suspension cultures, recapitulating the cellular architecture and molecular gradients in tissue. As such, they are popular for in vitro models in biological research. Classical imaging methodologies, like immunohistochemistry, are commonly used to examine the distribution of specific species within the spheroids. However, there is a need for an unbiased discovery-based methodology that would allow examination of protein/peptide distributions in 3D culture systems, without a need for prior knowledge of the analytes. We have developed a matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS)-based imaging approach to examine protein distributions in 3D cell culture models. Using colon carcinoma cell lines, we detect changes in the spatial distribution of proteins across 3D culture structures. To identify the protein species present, we are combining results from the MS/MS capabilities of MALDI-MS to sequence peptides in a de novo fashion and nanoflow liquid chromatography鈥搕andem mass spectrometry (nLC鈥揗S/MS) of homogenized cultures. As a proof-of-principle, we have identified cytochrome C and Histone H4 as two of the predominant protein species in the 3D colon carcinoma cultures.