title: Purification of centrosomes from mammalian cell lines.
abstract: Centrosomes act as the main microtubule-organizing centre of animal cells and play critical roles in the cell, such as mitotic spindle organization, cell polarity, and motility. They are composed of two barrel-shaped structures, the centrioles, surrounded by the pericentriolar matrix. In mammalian cells, the two centrioles differ structurally due to generational difference, the oldest one bearing appendages which allow the transient docking of the centriole at the plasma membrane in order to grow a primary cilium. Centrosome components are highly conserved throughout evolution and several pathologies have been associated with centrosomal defects. The understanding of such a complex organelle has therefore been a challenge for many researchers and has led to the development of centrosomal purification procedures to assess molecular composition, biological function, and structural organization of centrosomes. In this paper, we detail a step-by-step procedure to generate high yield of purified centrosome obtained from various mammalian cell lines.
authors.affiliation: Institut Curie, CNRS UMR144, Paris, France; Institut Curie, Orsay, France.
authors.forename: Delphine
authors.initials: D
authors.lastname: Gogendeau
authors.affiliation: Swiss Institute for Experimental Cancer Research (ISREC), School of Life Sciences, Swiss Federal Institute of Technology (EPFL), Lausanne, Switzerland.
authors.forename: Paul
authors.initials: P
authors.lastname: Guichard
authors.affiliation: Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Université Paris Sud, Gif sur Yvette, France.
authors.forename: Anne-Marie
authors.initials: AM
authors.lastname: Tassin
mesh_terms: D002458:Cell Fractionation / Q000379:methods; D002460:Cell Line; D018385:Centrosome; D006801:Humans
keywords: Centrioles; Centrosome isolation; Centrosomes; IIA1.6 cell line; KE37 cell line; RPE1 cell line