In this talk, I will present a single-cell mass spectrometry approach we have recently developed to characterize the metabolic composition of single embryonic cells in early-stage, 16-cell embryo of the South African clawed frog (Xenopus laevis), a favorite model in cell and developmental biology. We will discuss our approach to downscale traditional metabolomic workflows in mass spectrometry to single embryonic cells, specifically how we harvest single embryonic cells from the 16-cell embryo, efficiently extract metabolites, and use a custom-built capillary electrophoresis electrospray ionization system with a commercial time-of-flight mass spectrometer to obtain qualitative and quantitative information on cell-to-cell differences in the embryo. With high-efficiency separation, accurate mass measurements, and tandem mass spectrometry, we confidently identified an appreciable set of metabolites in the metabolome of embryonic cells. Relative quantitation revealed that certain cell types exhibit metabolic heterogeneity in 16-cell Xenopus embryos. Furthermore, in functional studies we found that certain metabolites were capable to altering the cell's normal developmental fate.
We anticipate that the adaptation of mass spectrometry to single embryonic cells holds exciting, new opportunities for cell and developmental biology as well as other fields where samples are limited in volume or mass. Funding information: This research is funded by National Institutes of Health Grant R21 GM114854 and The George Washington University Department of Chemistry Start-Up Funds.
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