We are developing a pulsed isotopic labeling strategy to rapidly examine the proteome of hypoxic versus normoxic cellular populations in spheroid tumor models. Spheroids grow outward in a radially symmetric fashion. An original nucleus of cells divides and proliferates in consecutively larger concentric spheres to form the structure, with the inner cells existing in lower oxygen concentrations compared to the outer cells. We are taking advantage of this growth pattern by sequentially adding pulsed SILAC (pSILAC) labels to the cell culture media (Light, Medium and Heavy isotopic labels). By labeling the different populations of cells in the spheroids with distinct isotopic labels, their origins within the structure can be discriminated by the mass spectrometer. We are exploring the protein differences in cells from the high versus low oxygen conditions with this approach.<br/>

Pulsed isotopic labeling of hypoxic and normoxic cellular populations

2019 Spring National Meeting

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