1. Proteome analysis of the culture environment supporting undifferentiated mouse embryonic stem and germ cell growth
- Author
-
Christine Carapito, Nicolas Buhr, Agnès Hovasse, Stéphane Viville, Alain Van Dorsselaer, Christine Schaeffer, Institut de génétique et biologie moléculaire et cellulaire (IGBMC), Université Louis Pasteur - Strasbourg I-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Spectrométrie de masse Bio-organique, UMR CNRS-ULP, Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Louis Pasteur - Strasbourg I, Laboratoire de Spectrométrie de Masse BioOrganique [Strasbourg] (LSMBO), Département Sciences Analytiques et Interactions Ioniques et Biomoléculaires (DSA-IPHC), Institut Pluridisciplinaire Hubert Curien (IPHC), Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Institut Pluridisciplinaire Hubert Curien (IPHC), and Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)
- Subjects
Proteomics ,Clinical Biochemistry ,Cell ,Bioinformatics ,Biochemistry ,Analytical Chemistry ,Mice ,0302 clinical medicine ,MESH: Animals ,Electrophoresis, Gel, Two-Dimensional ,MESH: Embryonic Stem Cells ,Cells, Cultured ,reproductive and urinary physiology ,0303 health sciences ,Ethical issues ,Culture environment ,MESH: Proteomics ,Reference Standards ,Cell biology ,medicine.anatomical_structure ,embryonic structures ,Proteome ,MESH: Pluripotent Stem Cells ,MESH: Reference Standards ,MESH: Gelatin ,biological phenomena, cell phenomena, and immunity ,Germ cell ,MESH: Cells, Cultured ,Pluripotent Stem Cells ,Biology ,Mouse embryonic fibroblast ,MESH: Coculture Techniques ,03 medical and health sciences ,MESH: Cell Proliferation ,medicine ,Animals ,Fibroblast ,MESH: Mice ,Embryonic Stem Cells ,Cell Proliferation ,030304 developmental biology ,urogenital system ,[SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology ,Fibroblasts ,MESH: Electrophoresis, Gel, Two-Dimensional ,Embryonic stem cell ,Coculture Techniques ,Culture Media ,MESH: Germ Cells ,Germ Cells ,MESH: Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,MESH: Fibroblasts ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,MESH: Culture Media ,Gelatin ,030217 neurology & neurosurgery - Abstract
The therapeutical interest of pluripotent cells and ethical issues related to the establishment of human embryonic stem cell (ESC) or embryonic germ cell (EGC) lines raise the understanding of the mechanism underlying pluripotency to a fundamental issue. Establishing a protein pluripotency signature for these cells can be complicated by the presence of unrelated proteins produced by the culture environment. Here, we have analyzed the environment supporting ESC and EGC growth, and established 2-D reference maps for each constituent present in this culture environment: mouse embryonic fibroblast feeder cells, culture medium (CM) and gelatin. The establishment of these reference maps is essential prior to the study of ESC and EGC specific proteomes. Indeed, these maps can be subtracted from ESC or EGC maps to allow focusing on spots specific for ESCs or EGCs. Our study led to the identification of 110 unique proteins from fibroblast feeder cells and 23 unique proteins from the CM, which represent major contaminants of ESC and EGC proteomes. For gelatin, no collagen-specific proteins were identified, most likely due to difficulties in resolution and low quantities. Furthermore, no differences were observed between naive and conditioned CM. Finally, we compared these reference maps to ESC 2-D gels and isolated 17 ESC specific spots. Among these spots, proteins that had already been identified in previous human and mouse ESC proteomes were identified but no apparent ESC-specific pluripotency marker could be identified. This work represents an essential step in furthering the knowledge of environmental factors supporting ESC and EGC growth.
- Published
- 2007