title: Rapid detection of apoptosis in mammalian cells by using intact cell MALDI mass spectrometry.
abstract: Detection of cell death has extensive applications and is of great commercial value. However, most current high-throughput cell viability assays cannot distinguish the two major forms of cell death: apoptosis and necrosis. Many apoptosis-specific detection methods exist but they are time consuming and labour intensive. In this work, we proposed a novel approach based on Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight Mass Spectrometry (MALDI-TOF-MS) for the specific detection of apoptosis in cultured mammalian cells. Buffer washed cells were directly mixed with a matrix solution and subsequently deposited onto the stainless steel target for MALDI analysis. The resulting mass spectrometric profiles were highly reproducible and can be used to reflect cell viability. Remarkably, the mass spectrometric profiles generated from apoptotic cells were distinct from those from either normal or necrotic cells. The apoptosis-specific features of the mass spectra were proportional to the percentage of apoptotic cells in the culture, but are independent of the drugs used to stimulate apoptosis. This is the first report on the utilization of intact cell MALDI mass spectrometry in detecting mammalian cell apoptosis, and can be used as a basis for the development of a reliable, fast, label-free and high-throughput method for detecting apoptotic cell death.
authors: Department of Chemistry and Biology, City University of Hong Kong, Kowloon, Hong Kong.
authors: Hongjuan
authors: H
authors: Dong
authors: Wei
authors: W
authors: Shen
authors: Myra Ting Wai
authors: MT
authors: Cheung
authors: Yimin
authors: Y
authors: Liang
authors: Hon Yeung
authors: HY
authors: Cheung
authors: Günter
authors: G
authors: Allmaier
authors: Oscar
authors: O
authors: Kin-Chung Au
authors: Yun Wah
authors: YW
authors: Lam
mesh_terms: D000818:Animals; D017209:Apoptosis; D002166:Camptothecin / Q000633:toxicity; D002460:Cell Line; D004224:Diterpenes / Q000633:toxicity; D004285:Dogs; D005047:Etoposide / Q000633:toxicity; D006801:Humans; D051379:Mice; D011419:Propidium / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D004224:Diterpenes; D011419:Propidium; C030419:andrographolide; D005047:Etoposide; D002166:Camptothecin