دورية أكاديمية
Subcellular fractionation of brain tumor stem cells.
| العنوان: | Subcellular fractionation of brain tumor stem cells. |
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| المؤلفون: | Sharanek A; Department of Cellular and Molecular Medicine and University of Ottawa Brain and Mind Research Institute, University of Ottawa, Ottawa, ON, Canada; Lady Davis Institute for Medical Research, Jewish General Hospital, Chemin de la Côte-Sainte-Catherine, Montréal, QC, Canada., Raco L; Lady Davis Institute for Medical Research, Jewish General Hospital, Chemin de la Côte-Sainte-Catherine, Montréal, QC, Canada; Division of Experimental Medicine, McGill University, Montréal, QC, Canada., Soleimani VD; Lady Davis Institute for Medical Research, Jewish General Hospital, Chemin de la Côte-Sainte-Catherine, Montréal, QC, Canada; Division of Experimental Medicine, McGill University, Montréal, QC, Canada; Department of Human Genetics, McGill University, Montréal, QC, Canada., Jahani-Asl A; Department of Cellular and Molecular Medicine and University of Ottawa Brain and Mind Research Institute, University of Ottawa, Ottawa, ON, Canada; Division of Experimental Medicine, McGill University, Montréal, QC, Canada. Electronic address: arezu.jahani@uottawa.ca. |
| المصدر: | Methods in cell biology [Methods Cell Biol] 2022; Vol. 170, pp. 47-58. Date of Electronic Publication: 2022 Apr 18. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Academic Press Country of Publication: United States NLM ID: 0373334 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 0091-679X (Print) Linking ISSN: 0091679X NLM ISO Abbreviation: Methods Cell Biol Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: New York, Academic Press |
| مواضيع طبية MeSH: | Neoplastic Stem Cells*/pathology , Proteomics*/methods, Animals ; Brain/metabolism ; Cell Fractionation/methods ; Cell Nucleus/metabolism ; Mammals/metabolism ; Proteome/metabolism ; Subcellular Fractions/metabolism |
| مستخلص: | Brain tumor stem cells (BTSCs) are a rare population of self-renewing stem cells that are cultured as spheres and are often slow growing compared to other mammalian cell lines. Analysis of BTSC proteome requires careful handling as well as techniques that can be applied to small quantities of cell material. Subcellular fractionation is a widely used technique to assess protein localization. Although proteins are often destined to a defined cell compartment via a signal peptide such as mitochondrial or nuclear localization signals, the recruitment of a protein from one compartment to another can occur as a result of post-translational modification and/or structural variations in response to intracellular and extracellular stimuli. These events assign different functions to a protein making the study of protein localization a useful approach for better understanding of its role in disease progression. Sequential centrifugation remains a simple and versatile fractionation method for proteomic analysis. It can also be applied for diverse downstream applications such as multi-omics using pure nuclear fractions or metabolomic studies on isolated mitochondria. In this chapter, we describe our optimized protocol for subcellular fractionation of BTSC spheres in which we use a commercially available kit with additional centrifugation steps. We provide details on BTSC maintenance and handling, fractionation protocol and evaluation of fraction purity. (Copyright © 2022 Elsevier Inc. All rights reserved.) |
| معلومات مُعتمدة: | Canada CIHR |
| فهرسة مساهمة: | Keywords: Brain tumor stem cells; Mitochondria; Stem cells; Subcellular fractionation |
| المشرفين على المادة: | 0 (Proteome) |
| تواريخ الأحداث: | Date Created: 20220710 Date Completed: 20220712 Latest Revision: 20220718 |
| رمز التحديث: | 20240628 |
| DOI: | 10.1016/bs.mcb.2022.02.006 |
| PMID: | 35811103 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 0091-679X |
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| DOI: | 10.1016/bs.mcb.2022.02.006 |