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Phenotypic and Molecular Characterization of Domestic Cat (Felis catus) Spermatogonial Stem Cells
In many mammalian species, surface markers have been used to obtain enriched populations of spermatogonial stem cells (SSCs) for assisted reproduction and other applications; however, little is known about the expression patterns of feline SSCs. In this study, we assessed expression of the SSC surfa...
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Published in: | Biology of reproduction 2016-07, Vol.95 (1), p.20-20 |
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creator | Powell, Robin H Galiguis, Jason Biancardi, Monica N Pope, C Earle Leibo, Stanley P Wang, Guoshun Gómez, Martha C |
description | In many mammalian species, surface markers have been used to obtain enriched populations of spermatogonial stem cells (SSCs) for assisted reproduction and other applications; however, little is known about the expression patterns of feline SSCs. In this study, we assessed expression of the SSC surface markers commonly used in other species, KIT, ITGA6, CD9, GFRalpha1, ADGRA3, and THY1, in addition to the less frequently used pluripotent markers TRA-1-60, TRA-1-81, SSEA-1, and SSEA-4 in SSCs of both prepubertal and adult domestic cats (Felis catus). To further characterize cat SSCs, we sorted cells using SSC-specific markers and evaluated the expression of the pluripotent transcription factors NANOG, POU5F1, and SOX2 and the proto-oncogene MYC within these populations. We concluded that SSC surface markers used in other mammalian species were not specific for identifying cat SSCs. However, the pluripotent markers we evaluated were more specific to cat spermatogonia, and the presence of SSEA-1 and SSEA-4 in fewer and primarily individual cells suggests that these two markers may be used for enrichment of cat SSCs. The expression of pluripotent transcription factors at mRNA level by single-stained cells positive for SSEA-4 and by dual-stained cells positive for both GFRalpha1 and SSEA-4 reflects the undifferentiated stage of cat SSCs. The absence of transcription factors in double-stained cells positive for only one marker implies the loss of the stem cell-like identity with the loss of either GFRalpha1 or SSEA-4. Further investigation is warranted to elucidate the biological characteristics of these spermatogonial subpopulations. |
doi_str_mv | 10.1095/biolreprod.115.134635 |
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In this study, we assessed expression of the SSC surface markers commonly used in other species, KIT, ITGA6, CD9, GFRalpha1, ADGRA3, and THY1, in addition to the less frequently used pluripotent markers TRA-1-60, TRA-1-81, SSEA-1, and SSEA-4 in SSCs of both prepubertal and adult domestic cats (Felis catus). To further characterize cat SSCs, we sorted cells using SSC-specific markers and evaluated the expression of the pluripotent transcription factors NANOG, POU5F1, and SOX2 and the proto-oncogene MYC within these populations. We concluded that SSC surface markers used in other mammalian species were not specific for identifying cat SSCs. However, the pluripotent markers we evaluated were more specific to cat spermatogonia, and the presence of SSEA-1 and SSEA-4 in fewer and primarily individual cells suggests that these two markers may be used for enrichment of cat SSCs. The expression of pluripotent transcription factors at mRNA level by single-stained cells positive for SSEA-4 and by dual-stained cells positive for both GFRalpha1 and SSEA-4 reflects the undifferentiated stage of cat SSCs. The absence of transcription factors in double-stained cells positive for only one marker implies the loss of the stem cell-like identity with the loss of either GFRalpha1 or SSEA-4. Further investigation is warranted to elucidate the biological characteristics of these spermatogonial subpopulations.</description><identifier>ISSN: 0006-3363</identifier><identifier>EISSN: 1529-7268</identifier><identifier>DOI: 10.1095/biolreprod.115.134635</identifier><identifier>PMID: 27281702</identifier><language>eng</language><publisher>United States</publisher><subject>Adult Germline Stem Cells - cytology ; Adult Germline Stem Cells - metabolism ; Animals ; Cats ; Cell Differentiation - physiology ; Integrin alpha6 - metabolism ; Lewis X Antigen - metabolism ; Male ; Nanog Homeobox Protein - metabolism ; Octamer Transcription Factor-3 - metabolism ; Proto-Oncogene Proteins c-kit - metabolism ; SOXB1 Transcription Factors - metabolism ; Spermatogonia - cytology ; Spermatogonia - metabolism ; Stage-Specific Embryonic Antigens - metabolism ; Tetraspanin-29 - metabolism</subject><ispartof>Biology of reproduction, 2016-07, Vol.95 (1), p.20-20</ispartof><rights>2016 by the Society for the Study of Reproduction, Inc.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c426t-38371d1abc6ec8ba0ab2343295d534f340a6d42f4fabc82205580a4cdc57617c3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,777,781,27905,27906</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/27281702$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Powell, Robin H</creatorcontrib><creatorcontrib>Galiguis, Jason</creatorcontrib><creatorcontrib>Biancardi, Monica N</creatorcontrib><creatorcontrib>Pope, C Earle</creatorcontrib><creatorcontrib>Leibo, Stanley P</creatorcontrib><creatorcontrib>Wang, Guoshun</creatorcontrib><creatorcontrib>Gómez, Martha C</creatorcontrib><title>Phenotypic and Molecular Characterization of Domestic Cat (Felis catus) Spermatogonial Stem Cells</title><title>Biology of reproduction</title><addtitle>Biol Reprod</addtitle><description>In many mammalian species, surface markers have been used to obtain enriched populations of spermatogonial stem cells (SSCs) for assisted reproduction and other applications; however, little is known about the expression patterns of feline SSCs. In this study, we assessed expression of the SSC surface markers commonly used in other species, KIT, ITGA6, CD9, GFRalpha1, ADGRA3, and THY1, in addition to the less frequently used pluripotent markers TRA-1-60, TRA-1-81, SSEA-1, and SSEA-4 in SSCs of both prepubertal and adult domestic cats (Felis catus). To further characterize cat SSCs, we sorted cells using SSC-specific markers and evaluated the expression of the pluripotent transcription factors NANOG, POU5F1, and SOX2 and the proto-oncogene MYC within these populations. We concluded that SSC surface markers used in other mammalian species were not specific for identifying cat SSCs. However, the pluripotent markers we evaluated were more specific to cat spermatogonia, and the presence of SSEA-1 and SSEA-4 in fewer and primarily individual cells suggests that these two markers may be used for enrichment of cat SSCs. The expression of pluripotent transcription factors at mRNA level by single-stained cells positive for SSEA-4 and by dual-stained cells positive for both GFRalpha1 and SSEA-4 reflects the undifferentiated stage of cat SSCs. The absence of transcription factors in double-stained cells positive for only one marker implies the loss of the stem cell-like identity with the loss of either GFRalpha1 or SSEA-4. Further investigation is warranted to elucidate the biological characteristics of these spermatogonial subpopulations.</description><subject>Adult Germline Stem Cells - cytology</subject><subject>Adult Germline Stem Cells - metabolism</subject><subject>Animals</subject><subject>Cats</subject><subject>Cell Differentiation - physiology</subject><subject>Integrin alpha6 - metabolism</subject><subject>Lewis X Antigen - metabolism</subject><subject>Male</subject><subject>Nanog Homeobox Protein - metabolism</subject><subject>Octamer Transcription Factor-3 - metabolism</subject><subject>Proto-Oncogene Proteins c-kit - metabolism</subject><subject>SOXB1 Transcription Factors - metabolism</subject><subject>Spermatogonia - cytology</subject><subject>Spermatogonia - metabolism</subject><subject>Stage-Specific Embryonic Antigens - metabolism</subject><subject>Tetraspanin-29 - metabolism</subject><issn>0006-3363</issn><issn>1529-7268</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><recordid>eNpFkMtOwzAQRS0EoqXwCSAvYZHid9IlChSQikAqrKOJ49CgJA62syhfj1ELLEazOXfm6iB0TsmckoW8LhvbOjM4W80plXPKheLyAE2pZIskZSo7RFNCiEo4V3yCTrz_IIQKzvgxmrCUZTQlbIrgZWN6G7ZDozH0FX6yrdFjCw7nG3Cgg3HNF4TG9tjW-NZ2xoeI5hDw5dK0jccawuiv8HowroNg323fQIvXwXQ4N23rT9FRDa03Z_s9Q2_Lu9f8IVk93z_mN6tEC6ZCwjOe0opCqZXRWQkESsZj3YWsJBc1FwRUJVgt6ohkjBEpMwJCV1qmiqaaz9Dl7m508jnGmkXXeB0bQG_s6AuaESUEF3FmSO5Q7az3ztTF4JoO3LagpPixW_zbLaLdYmc35i72L8ayM9Vf6lcn_wYEX3lA</recordid><startdate>20160701</startdate><enddate>20160701</enddate><creator>Powell, Robin H</creator><creator>Galiguis, Jason</creator><creator>Biancardi, Monica N</creator><creator>Pope, C Earle</creator><creator>Leibo, Stanley P</creator><creator>Wang, Guoshun</creator><creator>Gómez, Martha C</creator><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20160701</creationdate><title>Phenotypic and Molecular Characterization of Domestic Cat (Felis catus) Spermatogonial Stem Cells</title><author>Powell, Robin H ; Galiguis, Jason ; Biancardi, Monica N ; Pope, C Earle ; Leibo, Stanley P ; Wang, Guoshun ; Gómez, Martha C</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c426t-38371d1abc6ec8ba0ab2343295d534f340a6d42f4fabc82205580a4cdc57617c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Adult Germline Stem Cells - cytology</topic><topic>Adult Germline Stem Cells - metabolism</topic><topic>Animals</topic><topic>Cats</topic><topic>Cell Differentiation - physiology</topic><topic>Integrin alpha6 - metabolism</topic><topic>Lewis X Antigen - metabolism</topic><topic>Male</topic><topic>Nanog Homeobox Protein - metabolism</topic><topic>Octamer Transcription Factor-3 - metabolism</topic><topic>Proto-Oncogene Proteins c-kit - metabolism</topic><topic>SOXB1 Transcription Factors - metabolism</topic><topic>Spermatogonia - cytology</topic><topic>Spermatogonia - metabolism</topic><topic>Stage-Specific Embryonic Antigens - metabolism</topic><topic>Tetraspanin-29 - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Powell, Robin H</creatorcontrib><creatorcontrib>Galiguis, Jason</creatorcontrib><creatorcontrib>Biancardi, Monica N</creatorcontrib><creatorcontrib>Pope, C Earle</creatorcontrib><creatorcontrib>Leibo, Stanley P</creatorcontrib><creatorcontrib>Wang, Guoshun</creatorcontrib><creatorcontrib>Gómez, Martha C</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Biology of reproduction</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Powell, Robin H</au><au>Galiguis, Jason</au><au>Biancardi, Monica N</au><au>Pope, C Earle</au><au>Leibo, Stanley P</au><au>Wang, Guoshun</au><au>Gómez, Martha C</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Phenotypic and Molecular Characterization of Domestic Cat (Felis catus) Spermatogonial Stem Cells</atitle><jtitle>Biology of reproduction</jtitle><addtitle>Biol Reprod</addtitle><date>2016-07-01</date><risdate>2016</risdate><volume>95</volume><issue>1</issue><spage>20</spage><epage>20</epage><pages>20-20</pages><issn>0006-3363</issn><eissn>1529-7268</eissn><abstract>In many mammalian species, surface markers have been used to obtain enriched populations of spermatogonial stem cells (SSCs) for assisted reproduction and other applications; however, little is known about the expression patterns of feline SSCs. 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subjects | Adult Germline Stem Cells - cytology Adult Germline Stem Cells - metabolism Animals Cats Cell Differentiation - physiology Integrin alpha6 - metabolism Lewis X Antigen - metabolism Male Nanog Homeobox Protein - metabolism Octamer Transcription Factor-3 - metabolism Proto-Oncogene Proteins c-kit - metabolism SOXB1 Transcription Factors - metabolism Spermatogonia - cytology Spermatogonia - metabolism Stage-Specific Embryonic Antigens - metabolism Tetraspanin-29 - metabolism |
title | Phenotypic and Molecular Characterization of Domestic Cat (Felis catus) Spermatogonial Stem Cells |
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