Differing lectin binding profiles among human embryonic stem cells and derivatives aid in the isolation of neural progenitor cells
Human embryonic stem cells (hESCs) and their differentiated progeny allow for investigation of important changes/events during normal embryonic development. Currently most of the research is focused on proteinacous changes occurring as a result of differentiation of stem cells and little is known ab...
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| Veröffentlicht in: | PloS one 2011-08, Vol.6 (8), p.e23266-e23266 |
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| creator | Dodla, Mahesh C Young, Amber Venable, Alison Hasneen, Kowser Rao, Raj R Machacek, David W Stice, Steven L |
| description | Human embryonic stem cells (hESCs) and their differentiated progeny allow for investigation of important changes/events during normal embryonic development. Currently most of the research is focused on proteinacous changes occurring as a result of differentiation of stem cells and little is known about changes in cell surface glycosylation patterns. Identification of cell lineage specific glycans can help in understanding their role in maintenance, proliferation and differentiation. Furthermore, these glycans can serve as markers for isolation of homogenous populations of cells. Using a panel of eight biotinylated lectins, the glycan expression of hESCs, hESCs-derived human neural progenitors (hNP) cells, and hESCs-derived mesenchymal progenitor (hMP) cells was investigated. Our goal was to identify glycans that are unique for hNP cells and use the corresponding lectins for cell isolation. Flow cytometry and immunocytochemistry were used to determine expression and localization of glycans, respectively, in each cell type. These results show that the glycan expression changes upon differentiation of hESCs and is different for neural and mesenchymal lineage. For example, binding of PHA-L lectin is low in hESCs (14±4.4%) but significantly higher in differentiated hNP cells (99±0.4%) and hMP cells (90±3%). Three lectins: VVA, DBA and LTL have low binding in hESCs and hMP cells, but significantly higher binding in hNP cells. Finally, VVA lectin binding was used to isolate hNP cells from a mixed population of hESCs, hNP cells and hMP cells. This is the first report that compares glycan expression across these human stem cell lineages and identifies significant differences. Also, this is the first study that uses VVA lectin for isolation for human neural progenitor cells. |
| doi_str_mv | 10.1371/journal.pone.0023266 |
| format | Article |
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Currently most of the research is focused on proteinacous changes occurring as a result of differentiation of stem cells and little is known about changes in cell surface glycosylation patterns. Identification of cell lineage specific glycans can help in understanding their role in maintenance, proliferation and differentiation. Furthermore, these glycans can serve as markers for isolation of homogenous populations of cells. Using a panel of eight biotinylated lectins, the glycan expression of hESCs, hESCs-derived human neural progenitors (hNP) cells, and hESCs-derived mesenchymal progenitor (hMP) cells was investigated. Our goal was to identify glycans that are unique for hNP cells and use the corresponding lectins for cell isolation. Flow cytometry and immunocytochemistry were used to determine expression and localization of glycans, respectively, in each cell type. These results show that the glycan expression changes upon differentiation of hESCs and is different for neural and mesenchymal lineage. For example, binding of PHA-L lectin is low in hESCs (14±4.4%) but significantly higher in differentiated hNP cells (99±0.4%) and hMP cells (90±3%). Three lectins: VVA, DBA and LTL have low binding in hESCs and hMP cells, but significantly higher binding in hNP cells. Finally, VVA lectin binding was used to isolate hNP cells from a mixed population of hESCs, hNP cells and hMP cells. This is the first report that compares glycan expression across these human stem cell lineages and identifies significant differences. Also, this is the first study that uses VVA lectin for isolation for human neural progenitor cells.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0023266</identifier><identifier>PMID: 21850265</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Binding ; Biology ; Carbohydrates ; Cell Lineage ; Cell Separation - methods ; Cell surface ; Cells (biology) ; Cells, Cultured ; Chondroitin sulfate ; Cytometry ; Differentiation ; Embryo cells ; Embryogenesis ; Embryonic growth stage ; Embryonic stem cells ; Embryonic Stem Cells - cytology ; Embryonic Stem Cells - metabolism ; Embryos ; Flow Cytometry ; Glycan ; Glycosylation ; Humans ; Immunocytochemistry ; Immunohistochemistry ; Import regulations ; Laboratories ; Lectins ; Lectins - metabolism ; Localization ; Mesenchyme ; Nervous system ; Neural stem cells ; Neurogenesis ; Neurons - cytology ; Neurons - metabolism ; Polysaccharides ; Progenitor cells ; Progeny ; Spinal cord ; Stem cell research ; Stem cells ; Stem Cells - cytology ; Stem Cells - metabolism ; Studies ; Transplants & implants</subject><ispartof>PloS one, 2011-08, Vol.6 (8), p.e23266-e23266</ispartof><rights>COPYRIGHT 2011 Public Library of Science</rights><rights>2011 Dodla et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: https://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 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Currently most of the research is focused on proteinacous changes occurring as a result of differentiation of stem cells and little is known about changes in cell surface glycosylation patterns. Identification of cell lineage specific glycans can help in understanding their role in maintenance, proliferation and differentiation. Furthermore, these glycans can serve as markers for isolation of homogenous populations of cells. Using a panel of eight biotinylated lectins, the glycan expression of hESCs, hESCs-derived human neural progenitors (hNP) cells, and hESCs-derived mesenchymal progenitor (hMP) cells was investigated. Our goal was to identify glycans that are unique for hNP cells and use the corresponding lectins for cell isolation. Flow cytometry and immunocytochemistry were used to determine expression and localization of glycans, respectively, in each cell type. These results show that the glycan expression changes upon differentiation of hESCs and is different for neural and mesenchymal lineage. For example, binding of PHA-L lectin is low in hESCs (14±4.4%) but significantly higher in differentiated hNP cells (99±0.4%) and hMP cells (90±3%). Three lectins: VVA, DBA and LTL have low binding in hESCs and hMP cells, but significantly higher binding in hNP cells. Finally, VVA lectin binding was used to isolate hNP cells from a mixed population of hESCs, hNP cells and hMP cells. This is the first report that compares glycan expression across these human stem cell lineages and identifies significant differences. Also, this is the first study that uses VVA lectin for isolation for human neural progenitor cells.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>21850265</pmid><doi>10.1371/journal.pone.0023266</doi><tpages>e23266</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2011-08, Vol.6 (8), p.e23266-e23266 |
| issn | 1932-6203 1932-6203 |
| language | eng |
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| source | MEDLINE; DOAJ Directory of Open Access Journals; Public Library of Science (PLoS) Journals Open Access; EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Binding Biology Carbohydrates Cell Lineage Cell Separation - methods Cell surface Cells (biology) Cells, Cultured Chondroitin sulfate Cytometry Differentiation Embryo cells Embryogenesis Embryonic growth stage Embryonic stem cells Embryonic Stem Cells - cytology Embryonic Stem Cells - metabolism Embryos Flow Cytometry Glycan Glycosylation Humans Immunocytochemistry Immunohistochemistry Import regulations Laboratories Lectins Lectins - metabolism Localization Mesenchyme Nervous system Neural stem cells Neurogenesis Neurons - cytology Neurons - metabolism Polysaccharides Progenitor cells Progeny Spinal cord Stem cell research Stem cells Stem Cells - cytology Stem Cells - metabolism Studies Transplants & implants |
| title | Differing lectin binding profiles among human embryonic stem cells and derivatives aid in the isolation of neural progenitor cells |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-25T04%3A53%3A36IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Differing%20lectin%20binding%20profiles%20among%20human%20embryonic%20stem%20cells%20and%20derivatives%20aid%20in%20the%20isolation%20of%20neural%20progenitor%20cells&rft.jtitle=PloS%20one&rft.au=Dodla,%20Mahesh%20C&rft.date=2011-08-05&rft.volume=6&rft.issue=8&rft.spage=e23266&rft.epage=e23266&rft.pages=e23266-e23266&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0023266&rft_dat=%3Cgale_plos_%3EA476882969%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1307261464&rft_id=info:pmid/21850265&rft_galeid=A476882969&rft_doaj_id=oai_doaj_org_article_acc1ac667cf747b898b5697a82dc509c&rfr_iscdi=true |