Spermatogonial stem cells identified by molecular expression of PLZF, integrin β1 and reactivity to Dolichos biflorus agglutinin in alpaca adult testes
The identification system of spermatogonial stem cell (SSC) was established in alpaca using the molecular expression as well as the reactivity pattern to Dolichos biflorus agglutinin (DBA) by flow cytometry. Twenty‐four testicles with their epididymis were recovered from adult alpacas at the slaught...
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| Veröffentlicht in: | Andrologia 2019-07, Vol.51 (6), p.e13283-n/a |
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| creator | Valdivia, Martha Castañeda‐Zegarra, Sergio Lévano, Gloria Lazo, Jorge Reyes, Jhakelin Bravo, Zezé Santiani, Alexei Mujica, Fidel Ruíz, Jaime Gonzales, Gustavo F. |
| description | The identification system of spermatogonial stem cell (SSC) was established in alpaca using the molecular expression as well as the reactivity pattern to Dolichos biflorus agglutinin (DBA) by flow cytometry. Twenty‐four testicles with their epididymis were recovered from adult alpacas at the slaughterhouse of Huancavelica—Perú. Samples were transported to the Laboratory of Reproductive Physiology at Universidad Nacional Mayor de San Marcos. Testes were selected for our study when the progressive motility of epididymal spermatozoa (ESPM) was above 30%. Isolation of SSC was performed with two enzymatic digestions. Finally, sperm viability was evaluated by means of the trypan blue vital stain in spermatogonial round cells. Samples with more than 80% viability were selected. Isolated cells cultured for 2 days were used for identifying the presence of SSCs by the expression of integrin β1 (116 bp) and PLZF (206 bp) genes. Spermatogonia were classified according to the DBA reactivity. Spermatogonia with a strong positive to DBA (sDBA+) were classified as SSC (Mean ± SEM=4.44 ± 0.68%). Spermatogonia in early differentiation stages stained weakly positive with DBA (wDBA+) (Mean ± SEM=37.44 ± 3.07%) and differentiated round cells as DBA negative (Mean ± SEM=54.12 ± 3.18%). With the use of molecular and DBA markers, it is possible to identify easily the spermatogonial stem cells in alpaca. |
| doi_str_mv | 10.1111/and.13283 |
| format | Article |
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Twenty‐four testicles with their epididymis were recovered from adult alpacas at the slaughterhouse of Huancavelica—Perú. Samples were transported to the Laboratory of Reproductive Physiology at Universidad Nacional Mayor de San Marcos. Testes were selected for our study when the progressive motility of epididymal spermatozoa (ESPM) was above 30%. Isolation of SSC was performed with two enzymatic digestions. Finally, sperm viability was evaluated by means of the trypan blue vital stain in spermatogonial round cells. Samples with more than 80% viability were selected. Isolated cells cultured for 2 days were used for identifying the presence of SSCs by the expression of integrin β1 (116 bp) and PLZF (206 bp) genes. Spermatogonia were classified according to the DBA reactivity. Spermatogonia with a strong positive to DBA (sDBA+) were classified as SSC (Mean ± SEM=4.44 ± 0.68%). Spermatogonia in early differentiation stages stained weakly positive with DBA (wDBA+) (Mean ± SEM=37.44 ± 3.07%) and differentiated round cells as DBA negative (Mean ± SEM=54.12 ± 3.18%). With the use of molecular and DBA markers, it is possible to identify easily the spermatogonial stem cells in alpaca.</description><identifier>ISSN: 0303-4569</identifier><identifier>EISSN: 1439-0272</identifier><identifier>DOI: 10.1111/and.13283</identifier><identifier>PMID: 30957907</identifier><language>eng</language><publisher>Germany: Wiley Subscription Services, Inc</publisher><subject>Abattoirs ; Adult Germline Stem Cells - physiology ; alpaca ; Animals ; Biomarkers - analysis ; Biomarkers - metabolism ; Camelids, New World ; Cell Differentiation ; Cell Separation - methods ; Cell Separation - veterinary ; Cells, Cultured ; Conservation of Natural Resources ; DBA ; epididymal sperm mobility ; Epididymis ; Flow cytometry ; Flow Cytometry - methods ; Flow Cytometry - veterinary ; Insemination, Artificial ; Integrin beta1 - analysis ; Integrin beta1 - metabolism ; Male ; molecular markers ; Plant Lectins - chemistry ; Promyelocytic Leukemia Zinc Finger Protein - analysis ; Promyelocytic Leukemia Zinc Finger Protein - metabolism ; Spermatogonia ; Spermatogonia - physiology ; SSC ; Staining and Labeling - methods ; Staining and Labeling - veterinary ; Stem cells ; Testes ; Testis - cytology ; Testis - metabolism</subject><ispartof>Andrologia, 2019-07, Vol.51 (6), p.e13283-n/a</ispartof><rights>2019 Blackwell Verlag GmbH</rights><rights>2019 Blackwell Verlag GmbH.</rights><rights>Copyright © 2019 Blackwell Verlag GmbH</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3033-44fd9c97a20b0ddab858e4ba6b3b37f91c2ba66e71103110e7262bd33206daab3</citedby><cites>FETCH-LOGICAL-c3033-44fd9c97a20b0ddab858e4ba6b3b37f91c2ba66e71103110e7262bd33206daab3</cites><orcidid>0000-0001-9740-5490 ; 0000-0002-8301-5536 ; 0000-0002-8539-9262 ; 0000-0001-9107-3712 ; 0000-0001-9562-3099 ; 0000-0003-1611-2894</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Fand.13283$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fand.13283$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,27901,27902,45550,45551</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/30957907$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Valdivia, Martha</creatorcontrib><creatorcontrib>Castañeda‐Zegarra, Sergio</creatorcontrib><creatorcontrib>Lévano, Gloria</creatorcontrib><creatorcontrib>Lazo, Jorge</creatorcontrib><creatorcontrib>Reyes, Jhakelin</creatorcontrib><creatorcontrib>Bravo, Zezé</creatorcontrib><creatorcontrib>Santiani, Alexei</creatorcontrib><creatorcontrib>Mujica, Fidel</creatorcontrib><creatorcontrib>Ruíz, Jaime</creatorcontrib><creatorcontrib>Gonzales, Gustavo F.</creatorcontrib><title>Spermatogonial stem cells identified by molecular expression of PLZF, integrin β1 and reactivity to Dolichos biflorus agglutinin in alpaca adult testes</title><title>Andrologia</title><addtitle>Andrologia</addtitle><description>The identification system of spermatogonial stem cell (SSC) was established in alpaca using the molecular expression as well as the reactivity pattern to Dolichos biflorus agglutinin (DBA) by flow cytometry. Twenty‐four testicles with their epididymis were recovered from adult alpacas at the slaughterhouse of Huancavelica—Perú. Samples were transported to the Laboratory of Reproductive Physiology at Universidad Nacional Mayor de San Marcos. Testes were selected for our study when the progressive motility of epididymal spermatozoa (ESPM) was above 30%. Isolation of SSC was performed with two enzymatic digestions. Finally, sperm viability was evaluated by means of the trypan blue vital stain in spermatogonial round cells. Samples with more than 80% viability were selected. Isolated cells cultured for 2 days were used for identifying the presence of SSCs by the expression of integrin β1 (116 bp) and PLZF (206 bp) genes. Spermatogonia were classified according to the DBA reactivity. Spermatogonia with a strong positive to DBA (sDBA+) were classified as SSC (Mean ± SEM=4.44 ± 0.68%). Spermatogonia in early differentiation stages stained weakly positive with DBA (wDBA+) (Mean ± SEM=37.44 ± 3.07%) and differentiated round cells as DBA negative (Mean ± SEM=54.12 ± 3.18%). With the use of molecular and DBA markers, it is possible to identify easily the spermatogonial stem cells in alpaca.</description><subject>Abattoirs</subject><subject>Adult Germline Stem Cells - physiology</subject><subject>alpaca</subject><subject>Animals</subject><subject>Biomarkers - analysis</subject><subject>Biomarkers - metabolism</subject><subject>Camelids, New World</subject><subject>Cell Differentiation</subject><subject>Cell Separation - methods</subject><subject>Cell Separation - veterinary</subject><subject>Cells, Cultured</subject><subject>Conservation of Natural Resources</subject><subject>DBA</subject><subject>epididymal sperm mobility</subject><subject>Epididymis</subject><subject>Flow cytometry</subject><subject>Flow Cytometry - methods</subject><subject>Flow Cytometry - veterinary</subject><subject>Insemination, Artificial</subject><subject>Integrin beta1 - analysis</subject><subject>Integrin beta1 - metabolism</subject><subject>Male</subject><subject>molecular markers</subject><subject>Plant Lectins - chemistry</subject><subject>Promyelocytic Leukemia Zinc Finger Protein - analysis</subject><subject>Promyelocytic Leukemia Zinc Finger Protein - metabolism</subject><subject>Spermatogonia</subject><subject>Spermatogonia - physiology</subject><subject>SSC</subject><subject>Staining and Labeling - methods</subject><subject>Staining and Labeling - veterinary</subject><subject>Stem cells</subject><subject>Testes</subject><subject>Testis - cytology</subject><subject>Testis - metabolism</subject><issn>0303-4569</issn><issn>1439-0272</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kd9qFDEUxoModqm98AUk4I1Ct82_mexcltZqYamCeuPNkD9n1pRMMiYZ7b6Jz-GD-Exmu9WLgoeEEPjly3fOh9BzSk5orVMV7AnlbMUfoQUVvFsSJtljtCCc8KVo2u4AHeV8Q2qJRkohnqIDTrpGdkQu0M-PE6RRlbiJwSmPc4ERG_A-Y2chFDc4sFhv8Rg9mNmrhOF2SpCziwHHAX9Yf7k8xi4U2CQX8O9fFFdHOIEyxX13ZYtLxBfRO_M1Zqzd4GOaM1abjZ-LC_VJXcpPyiis7OwLLlBN5GfoyaB8hqP78xB9vnzz6fzdcv3-7dX52Xppan-1QTHYznRSMaKJtUqvmhUIrVrNNZdDRw2rlxYkpYTXDZK1TFvOGWmtUpofold73SnFb3P9uh9d3g1ABYhz7hkjjaBENryiLx-gN3FOobqrFOctF6zdUa_3lEkx5wRDPyU3qrTtKel3ifV1Pv1dYpV9ca846xHsP_JvPhU43QM_nIft_5X6s-uLveQf0ZWh3g</recordid><startdate>201907</startdate><enddate>201907</enddate><creator>Valdivia, Martha</creator><creator>Castañeda‐Zegarra, Sergio</creator><creator>Lévano, Gloria</creator><creator>Lazo, Jorge</creator><creator>Reyes, Jhakelin</creator><creator>Bravo, Zezé</creator><creator>Santiani, Alexei</creator><creator>Mujica, Fidel</creator><creator>Ruíz, Jaime</creator><creator>Gonzales, Gustavo F.</creator><general>Wiley Subscription Services, Inc</general><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>7TM</scope><scope>K9.</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0001-9740-5490</orcidid><orcidid>https://orcid.org/0000-0002-8301-5536</orcidid><orcidid>https://orcid.org/0000-0002-8539-9262</orcidid><orcidid>https://orcid.org/0000-0001-9107-3712</orcidid><orcidid>https://orcid.org/0000-0001-9562-3099</orcidid><orcidid>https://orcid.org/0000-0003-1611-2894</orcidid></search><sort><creationdate>201907</creationdate><title>Spermatogonial stem cells identified by molecular expression of PLZF, integrin β1 and reactivity to Dolichos biflorus agglutinin in alpaca adult testes</title><author>Valdivia, Martha ; Castañeda‐Zegarra, Sergio ; Lévano, Gloria ; Lazo, Jorge ; Reyes, Jhakelin ; Bravo, Zezé ; Santiani, Alexei ; Mujica, Fidel ; Ruíz, Jaime ; Gonzales, Gustavo F.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3033-44fd9c97a20b0ddab858e4ba6b3b37f91c2ba66e71103110e7262bd33206daab3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2019</creationdate><topic>Abattoirs</topic><topic>Adult Germline Stem Cells - physiology</topic><topic>alpaca</topic><topic>Animals</topic><topic>Biomarkers - analysis</topic><topic>Biomarkers - metabolism</topic><topic>Camelids, New World</topic><topic>Cell Differentiation</topic><topic>Cell Separation - methods</topic><topic>Cell Separation - veterinary</topic><topic>Cells, Cultured</topic><topic>Conservation of Natural Resources</topic><topic>DBA</topic><topic>epididymal sperm mobility</topic><topic>Epididymis</topic><topic>Flow cytometry</topic><topic>Flow Cytometry - methods</topic><topic>Flow Cytometry - veterinary</topic><topic>Insemination, Artificial</topic><topic>Integrin beta1 - analysis</topic><topic>Integrin beta1 - metabolism</topic><topic>Male</topic><topic>molecular markers</topic><topic>Plant Lectins - chemistry</topic><topic>Promyelocytic Leukemia Zinc Finger Protein - analysis</topic><topic>Promyelocytic Leukemia Zinc Finger Protein - metabolism</topic><topic>Spermatogonia</topic><topic>Spermatogonia - physiology</topic><topic>SSC</topic><topic>Staining and Labeling - methods</topic><topic>Staining and Labeling - veterinary</topic><topic>Stem cells</topic><topic>Testes</topic><topic>Testis - cytology</topic><topic>Testis - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Valdivia, Martha</creatorcontrib><creatorcontrib>Castañeda‐Zegarra, Sergio</creatorcontrib><creatorcontrib>Lévano, Gloria</creatorcontrib><creatorcontrib>Lazo, Jorge</creatorcontrib><creatorcontrib>Reyes, Jhakelin</creatorcontrib><creatorcontrib>Bravo, Zezé</creatorcontrib><creatorcontrib>Santiani, Alexei</creatorcontrib><creatorcontrib>Mujica, Fidel</creatorcontrib><creatorcontrib>Ruíz, Jaime</creatorcontrib><creatorcontrib>Gonzales, Gustavo F.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>MEDLINE - Academic</collection><jtitle>Andrologia</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Valdivia, Martha</au><au>Castañeda‐Zegarra, Sergio</au><au>Lévano, Gloria</au><au>Lazo, Jorge</au><au>Reyes, Jhakelin</au><au>Bravo, Zezé</au><au>Santiani, Alexei</au><au>Mujica, Fidel</au><au>Ruíz, Jaime</au><au>Gonzales, Gustavo F.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Spermatogonial stem cells identified by molecular expression of PLZF, integrin β1 and reactivity to Dolichos biflorus agglutinin in alpaca adult testes</atitle><jtitle>Andrologia</jtitle><addtitle>Andrologia</addtitle><date>2019-07</date><risdate>2019</risdate><volume>51</volume><issue>6</issue><spage>e13283</spage><epage>n/a</epage><pages>e13283-n/a</pages><issn>0303-4569</issn><eissn>1439-0272</eissn><abstract>The identification system of spermatogonial stem cell (SSC) was established in alpaca using the molecular expression as well as the reactivity pattern to Dolichos biflorus agglutinin (DBA) by flow cytometry. Twenty‐four testicles with their epididymis were recovered from adult alpacas at the slaughterhouse of Huancavelica—Perú. Samples were transported to the Laboratory of Reproductive Physiology at Universidad Nacional Mayor de San Marcos. Testes were selected for our study when the progressive motility of epididymal spermatozoa (ESPM) was above 30%. Isolation of SSC was performed with two enzymatic digestions. Finally, sperm viability was evaluated by means of the trypan blue vital stain in spermatogonial round cells. Samples with more than 80% viability were selected. Isolated cells cultured for 2 days were used for identifying the presence of SSCs by the expression of integrin β1 (116 bp) and PLZF (206 bp) genes. Spermatogonia were classified according to the DBA reactivity. Spermatogonia with a strong positive to DBA (sDBA+) were classified as SSC (Mean ± SEM=4.44 ± 0.68%). Spermatogonia in early differentiation stages stained weakly positive with DBA (wDBA+) (Mean ± SEM=37.44 ± 3.07%) and differentiated round cells as DBA negative (Mean ± SEM=54.12 ± 3.18%). With the use of molecular and DBA markers, it is possible to identify easily the spermatogonial stem cells in alpaca.</abstract><cop>Germany</cop><pub>Wiley Subscription Services, Inc</pub><pmid>30957907</pmid><doi>10.1111/and.13283</doi><tpages>8</tpages><orcidid>https://orcid.org/0000-0001-9740-5490</orcidid><orcidid>https://orcid.org/0000-0002-8301-5536</orcidid><orcidid>https://orcid.org/0000-0002-8539-9262</orcidid><orcidid>https://orcid.org/0000-0001-9107-3712</orcidid><orcidid>https://orcid.org/0000-0001-9562-3099</orcidid><orcidid>https://orcid.org/0000-0003-1611-2894</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Abattoirs Adult Germline Stem Cells - physiology alpaca Animals Biomarkers - analysis Biomarkers - metabolism Camelids, New World Cell Differentiation Cell Separation - methods Cell Separation - veterinary Cells, Cultured Conservation of Natural Resources DBA epididymal sperm mobility Epididymis Flow cytometry Flow Cytometry - methods Flow Cytometry - veterinary Insemination, Artificial Integrin beta1 - analysis Integrin beta1 - metabolism Male molecular markers Plant Lectins - chemistry Promyelocytic Leukemia Zinc Finger Protein - analysis Promyelocytic Leukemia Zinc Finger Protein - metabolism Spermatogonia Spermatogonia - physiology SSC Staining and Labeling - methods Staining and Labeling - veterinary Stem cells Testes Testis - cytology Testis - metabolism |
| title | Spermatogonial stem cells identified by molecular expression of PLZF, integrin β1 and reactivity to Dolichos biflorus agglutinin in alpaca adult testes |
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