A mass spectrometry-based workflow for the proteomic analysis of in vitro cultured cell subsets isolated by means of laser capture microdissection

摘要: This paper describes a microproteomic workflow that is useful for simultaneously identifying and quantifying proteins from minimal number of morphotypically heterogeneous cultured adherent cells. The analytical strategy makes use laser capture microdissection, an effective means harvesting pure cell populations, label-free mass spectrometry. We optimised the with particular reference to fixation which crucial successful laser-based microdissection also downstream molecular studies. In addition, we defined minimum cells be isolated analysed satisfactory proteome coverage. To set up this workflow, choose human monocyte-derived macrophages spontaneously differentiated in vitro. These cells, under our culture conditions, show distinct morphotypes, reminiscent heterogeneity observed tissues various homeostatic pathological states, e.g. atherosclerosis. may provide new insights into biology pathology culture, particularly when other selection approaches are not suitable.