{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2024,1,17]],"date-time":"2024-01-17T03:01:40Z","timestamp":1705460500475},"reference-count":64,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2013,2,1]],"date-time":"2013-02-01T00:00:00Z","timestamp":1359676800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["CP Stem Cell Biology"],"published-print":{"date-parts":[[2013,2]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>This unit describes two basic protocols for the detection of the proliferation of neural stem cells (NSC). The first one addresses cell proliferation in cultures, starting with primary cell cultures isolated from the mouse subventricular zone (SVZ), in which SVZ\u2010derived NSC are kept in culture as neurospheres. By using this culture system, we are able to study different stages of adult neurogenesis, such as proliferation, differentiation, migration, and survival. Thus, in the first basic protocol, we describe two different techniques to evaluate cell proliferation based on EdU incorporation: (a) immunocytochemistry and (b) flow cytometry. EdU, a new thymidine analog, which is detected by a reproducible and sensitive method based on click chemistry, does not require DNA denaturation, as is the case with BrdU. Thus, co\u2010labeling of EdU with other specific antibodies of extracellular or intracellular targets, as well as other DNA dyes, is possible. In the second basic protocol, we describe an in vivo assay to evaluate proliferation of NSC in the dentate gyrus of hippocampus of adult mice, by both BrdU and EdU detection. With this approach, it is also possible to study different stages of adult neurogenesis, by co\u2010labeling thymidine analogs with other specific markers, such as doublecortin (DCX) or neuronal nuclei protein (NeuN). <jats:italic>Curr. Protoc. Stem Cell Biol<\/jats:italic>. 24:2D.14.1\u20102D.14.24. \u00a9 2013 by John Wiley &amp; Sons, Inc.<\/jats:p>","DOI":"10.1002\/9780470151808.sc02d14s24","type":"journal-article","created":{"date-parts":[[2013,2,13]],"date-time":"2013-02-13T13:01:27Z","timestamp":1360760487000},"source":"Crossref","is-referenced-by-count":12,"title":["Evaluation of Proliferation of Neural Stem Cells In Vitro and In Vivo"],"prefix":"10.1002","volume":"24","author":[{"given":"Maria In\u00eas","family":"Morte","sequence":"first","affiliation":[{"name":"Centre for Neuroscience and Cell Biology, Neuroendocrinology and Neurogenesis Group  University of Coimbra Coimbra, Portugal"},{"name":"These authors contributed equally to this work"}]},{"given":"Bruno P.","family":"Carreira","sequence":"additional","affiliation":[{"name":"Centre for Neuroscience and Cell Biology, Neuroendocrinology and Neurogenesis Group  University of Coimbra Coimbra, Portugal"},{"name":"These authors contributed equally to this work"}]},{"given":"Vanessa","family":"Machado","sequence":"additional","affiliation":[{"name":"Centre for Neuroscience and Cell Biology, Neuroendocrinology and Neurogenesis Group  University of Coimbra Coimbra, Portugal"},{"name":"Regenerative Medicine Program, Department of Biomedical Sciences and Medicine  University of Algarve Faro, Portugal"},{"name":"IBB\u2010Institute for Biotechnology and Bioengineering, Centre of Molecular and Structural Biomedicine  University of Algarve Faro, Portugal"}]},{"given":"An\u00e1lia","family":"Carmo","sequence":"additional","affiliation":[{"name":"Centre for Neuroscience and Cell Biology, Cellular Immunology and Oncobiology Group  University of Coimbra Coimbra, Portugal"}]},{"given":"Isabel","family":"Nunes\u2010Correia","sequence":"additional","affiliation":[{"name":"Centre for Neuroscience and Cell Biology, Flow Cytometry Unit  University of Coimbra Coimbra, Portugal"}]},{"given":"Caetana M.","family":"Carvalho","sequence":"additional","affiliation":[{"name":"Centre for Neuroscience and Cell Biology, Neuroendocrinology and Neurogenesis Group  University of Coimbra Coimbra, Portugal"}]},{"given":"In\u00eas M.","family":"Ara\u00fajo","sequence":"additional","affiliation":[{"name":"Centre for Neuroscience and Cell Biology, Neuroendocrinology and Neurogenesis Group  University of Coimbra Coimbra, Portugal"},{"name":"Regenerative Medicine Program, Department of Biomedical Sciences and Medicine  University of Algarve Faro, Portugal"},{"name":"IBB\u2010Institute for Biotechnology and Bioengineering, Centre of Molecular and Structural Biomedicine  University of Algarve Faro, Portugal"}]}],"member":"311","published-online":{"date-parts":[[2013,2]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1634\/stemcells.2008-0056"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1002\/cne.901370404"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1002\/cne.901240303"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0896-6273(04)00111-4"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.23-32-10411.2003"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/0014-5793(77)81006-5"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1002\/cne.10874"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0166-2236(00)01725-2"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1038\/88158"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0960-9822(02)00771-6"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.1002\/stem.444"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1002\/0471142735.im1400s96"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1242\/dev.061663"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1440-1681.2007.04609.x"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.1159\/000057572"},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.17-13-05046.1997"},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/0955-0674(94)90054-X"},{"key":"e_1_2_10_19_1","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.1031-07.2007"},{"key":"e_1_2_10_20_1","doi-asserted-by":"publisher","DOI":"10.2174\/138920107780906540"},{"key":"e_1_2_10_21_1","doi-asserted-by":"publisher","DOI":"10.1038\/3305"},{"key":"e_1_2_10_22_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.133.4.1710"},{"key":"e_1_2_10_23_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.97.4.1823"},{"key":"e_1_2_10_24_1","first-page":"95","article-title":"Characterization of oligodendrocyte lineage precursor cells in the mouse cerebral cortex: A confocal microscopy approach to demyelinating diseases","volume":"115","author":"Girolamo F.","year":"2010","journal-title":"Ital. 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