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Supplementation of culture medium with knockout serum replacement improves the survival of bovine secondary follicles when compared with other protein sources during in vitro culture
The present study evaluated the effect of knockout serum replacement (KSR), fetal bovine serum (FBS) and bovine serum albumin (BSA) on the viability and growth of bovine secondary follicles cultured in vitro for 12 days. To this end, secondary follicles were isolated (185-202 μm) and cultured in vit...
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Published in: | Zygote (Cambridge) 2020-02, Vol.28 (1), p.32-36 |
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creator | Gomes, D S Aragão, L B Lima Neto, M F Barroso, P A A Paulino, L R F M Silva, B R Souza, A L P Vasconcelos, G L Silva, A W B Silva, J R V |
description | The present study evaluated the effect of knockout serum replacement (KSR), fetal bovine serum (FBS) and bovine serum albumin (BSA) on the viability and growth of bovine secondary follicles cultured in vitro for 12 days. To this end, secondary follicles were isolated (185-202 μm) and cultured in vitro in TCM-199+ medium supplemented with KSR (5% and 10%), FBS (5% and 10%) or BSA (3 mg/ml) at 38.5°C with 5% CO2 in air. Follicular diameters were evaluated on days 0, 4, 8 and 12. After 12 days of culture, follicular survival analysis was performing by using calcein-AM and ethidium homodimer. Before and after culture, follicles were fixed in paraformaldehyde for histological evaluation. Follicular diameter at different days of culture were compared using the Kruskal-Wallis test, while the percentages of viable follicles were analyzed by chi-squared test (P < 0.05). Results showed that follicles cultured in the presence of KSR at both concentrations presented higher follicular survival rates than those cultured in control medium alone or supplemented with FBS or BSA. Conversely, the presence of KSR, BSA or FBS did not increase follicular diameter after 12 days of culture. Histology analysis showed that, among the tested treatments, follicles cultured in the presence of KSR had preserved rounded oocytes, juxtaposed granulosa cells and intact basal membrane. In conclusion, supplementation of culture medium with KSR increases the follicular survival of bovine secondary follicles cultured in vitro. |
doi_str_mv | 10.1017/S0967199419000583 |
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To this end, secondary follicles were isolated (185-202 μm) and cultured in vitro in TCM-199+ medium supplemented with KSR (5% and 10%), FBS (5% and 10%) or BSA (3 mg/ml) at 38.5°C with 5% CO2 in air. Follicular diameters were evaluated on days 0, 4, 8 and 12. After 12 days of culture, follicular survival analysis was performing by using calcein-AM and ethidium homodimer. Before and after culture, follicles were fixed in paraformaldehyde for histological evaluation. Follicular diameter at different days of culture were compared using the Kruskal-Wallis test, while the percentages of viable follicles were analyzed by chi-squared test (P < 0.05). Results showed that follicles cultured in the presence of KSR at both concentrations presented higher follicular survival rates than those cultured in control medium alone or supplemented with FBS or BSA. Conversely, the presence of KSR, BSA or FBS did not increase follicular diameter after 12 days of culture. Histology analysis showed that, among the tested treatments, follicles cultured in the presence of KSR had preserved rounded oocytes, juxtaposed granulosa cells and intact basal membrane. In conclusion, supplementation of culture medium with KSR increases the follicular survival of bovine secondary follicles cultured in vitro.</description><identifier>ISSN: 0967-1994</identifier><identifier>EISSN: 1469-8730</identifier><identifier>DOI: 10.1017/S0967199419000583</identifier><identifier>PMID: 31601279</identifier><language>eng</language><publisher>England: Cambridge University Press</publisher><subject>Bovine serum albumin ; Calcein ; Carbon dioxide ; Cell culture ; Chi-square test ; Embryos ; Fetal calf serum ; Follicles ; Gametocytes ; Granulosa cells ; Histology ; Laboratories ; Morphology ; Oocytes ; Protein sources ; Proteins ; Serum albumin ; Statistical tests ; Supplements ; Survival ; Variance analysis</subject><ispartof>Zygote (Cambridge), 2020-02, Vol.28 (1), p.32-36</ispartof><rights>Cambridge University Press 2019</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c244t-f07ae9865ae7933d62809b1a6d0279809bd9d7096c0c4a82d697725d8db1da053</citedby><cites>FETCH-LOGICAL-c244t-f07ae9865ae7933d62809b1a6d0279809bd9d7096c0c4a82d697725d8db1da053</cites><orcidid>0000-0002-5970-6177</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>315,786,790,27957,27958</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/31601279$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Gomes, D S</creatorcontrib><creatorcontrib>Aragão, L B</creatorcontrib><creatorcontrib>Lima Neto, M F</creatorcontrib><creatorcontrib>Barroso, P A A</creatorcontrib><creatorcontrib>Paulino, L R F M</creatorcontrib><creatorcontrib>Silva, B R</creatorcontrib><creatorcontrib>Souza, A L P</creatorcontrib><creatorcontrib>Vasconcelos, G L</creatorcontrib><creatorcontrib>Silva, A W B</creatorcontrib><creatorcontrib>Silva, J R V</creatorcontrib><title>Supplementation of culture medium with knockout serum replacement improves the survival of bovine secondary follicles when compared with other protein sources during in vitro culture</title><title>Zygote (Cambridge)</title><addtitle>Zygote</addtitle><description>The present study evaluated the effect of knockout serum replacement (KSR), fetal bovine serum (FBS) and bovine serum albumin (BSA) on the viability and growth of bovine secondary follicles cultured in vitro for 12 days. To this end, secondary follicles were isolated (185-202 μm) and cultured in vitro in TCM-199+ medium supplemented with KSR (5% and 10%), FBS (5% and 10%) or BSA (3 mg/ml) at 38.5°C with 5% CO2 in air. Follicular diameters were evaluated on days 0, 4, 8 and 12. After 12 days of culture, follicular survival analysis was performing by using calcein-AM and ethidium homodimer. Before and after culture, follicles were fixed in paraformaldehyde for histological evaluation. Follicular diameter at different days of culture were compared using the Kruskal-Wallis test, while the percentages of viable follicles were analyzed by chi-squared test (P < 0.05). Results showed that follicles cultured in the presence of KSR at both concentrations presented higher follicular survival rates than those cultured in control medium alone or supplemented with FBS or BSA. Conversely, the presence of KSR, BSA or FBS did not increase follicular diameter after 12 days of culture. Histology analysis showed that, among the tested treatments, follicles cultured in the presence of KSR had preserved rounded oocytes, juxtaposed granulosa cells and intact basal membrane. In conclusion, supplementation of culture medium with KSR increases the follicular survival of bovine secondary follicles cultured in vitro.</description><subject>Bovine serum albumin</subject><subject>Calcein</subject><subject>Carbon dioxide</subject><subject>Cell culture</subject><subject>Chi-square test</subject><subject>Embryos</subject><subject>Fetal calf serum</subject><subject>Follicles</subject><subject>Gametocytes</subject><subject>Granulosa cells</subject><subject>Histology</subject><subject>Laboratories</subject><subject>Morphology</subject><subject>Oocytes</subject><subject>Protein sources</subject><subject>Proteins</subject><subject>Serum albumin</subject><subject>Statistical tests</subject><subject>Supplements</subject><subject>Survival</subject><subject>Variance analysis</subject><issn>0967-1994</issn><issn>1469-8730</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><recordid>eNplUctu1TAQtRCI3l74ADbIEhs2AT-SOF6iqlCkSiwK68jXnnDdOnbwIxU_xvfhcFsWsLI1c86ZM3MQekXJO0qoeH9DZC-olC2VhJBu4E_Qjra9bAbByVO029rN1j9D5yndVowQsn2OzjjtCWVC7tCvm7IsDmbwWWUbPA4T1sXlEgHPYGyZ8b3NR3zng74LJeMEsdYiLE7pPzRs5yWGFRLOR8CpxNWuym06h7BaX0uggzcq_sRTcM5qV6H3R_BYh3lREcxpQqj0iKtUButxCiXqCjQlWv8d18pqcwyP3l6gZ5NyCV4-vHv07ePl14ur5vrLp88XH64bzdo2NxMRCuTQdwqE5Nz0bCDyQFVvSN1--xtpRD2TJrpVAzO9FIJ1ZjAHahTp-B69PelWXz8KpDzONmlwTnkIJY2Mk460nPVDhb75B3pbd_DV3cha0rKODtXDHtETSseQUoRpXKKd63FGSsYt1PG_UCvn9YNyOdRM_jIeU-S_ASwOoWA</recordid><startdate>202002</startdate><enddate>202002</enddate><creator>Gomes, D S</creator><creator>Aragão, L B</creator><creator>Lima Neto, M F</creator><creator>Barroso, P A A</creator><creator>Paulino, L R F M</creator><creator>Silva, B R</creator><creator>Souza, A L P</creator><creator>Vasconcelos, G L</creator><creator>Silva, A W B</creator><creator>Silva, J R V</creator><general>Cambridge University Press</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7P</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0002-5970-6177</orcidid></search><sort><creationdate>202002</creationdate><title>Supplementation of culture medium with knockout serum replacement improves the survival of bovine secondary follicles when compared with other protein sources during in vitro culture</title><author>Gomes, D S ; Aragão, L B ; Lima Neto, M F ; Barroso, P A A ; Paulino, L R F M ; Silva, B R ; Souza, A L P ; Vasconcelos, G L ; Silva, A W B ; Silva, J R V</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c244t-f07ae9865ae7933d62809b1a6d0279809bd9d7096c0c4a82d697725d8db1da053</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>Bovine serum albumin</topic><topic>Calcein</topic><topic>Carbon dioxide</topic><topic>Cell culture</topic><topic>Chi-square test</topic><topic>Embryos</topic><topic>Fetal calf serum</topic><topic>Follicles</topic><topic>Gametocytes</topic><topic>Granulosa cells</topic><topic>Histology</topic><topic>Laboratories</topic><topic>Morphology</topic><topic>Oocytes</topic><topic>Protein sources</topic><topic>Proteins</topic><topic>Serum albumin</topic><topic>Statistical tests</topic><topic>Supplements</topic><topic>Survival</topic><topic>Variance analysis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Gomes, D S</creatorcontrib><creatorcontrib>Aragão, L B</creatorcontrib><creatorcontrib>Lima Neto, M F</creatorcontrib><creatorcontrib>Barroso, P A A</creatorcontrib><creatorcontrib>Paulino, L R F M</creatorcontrib><creatorcontrib>Silva, B R</creatorcontrib><creatorcontrib>Souza, A L P</creatorcontrib><creatorcontrib>Vasconcelos, G L</creatorcontrib><creatorcontrib>Silva, A W B</creatorcontrib><creatorcontrib>Silva, J R V</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Agricultural Science Collection</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni)</collection><collection>ProQuest Central</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Biological Sciences</collection><collection>Agriculture Science Database</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>PML(ProQuest Medical Library)</collection><collection>Biological Science Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>MEDLINE - Academic</collection><jtitle>Zygote (Cambridge)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Gomes, D S</au><au>Aragão, L B</au><au>Lima Neto, M F</au><au>Barroso, P A A</au><au>Paulino, L R F M</au><au>Silva, B R</au><au>Souza, A L P</au><au>Vasconcelos, G L</au><au>Silva, A W B</au><au>Silva, J R V</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Supplementation of culture medium with knockout serum replacement improves the survival of bovine secondary follicles when compared with other protein sources during in vitro culture</atitle><jtitle>Zygote (Cambridge)</jtitle><addtitle>Zygote</addtitle><date>2020-02</date><risdate>2020</risdate><volume>28</volume><issue>1</issue><spage>32</spage><epage>36</epage><pages>32-36</pages><issn>0967-1994</issn><eissn>1469-8730</eissn><notes>ObjectType-Article-1</notes><notes>SourceType-Scholarly Journals-1</notes><notes>ObjectType-Feature-2</notes><notes>content type line 23</notes><abstract>The present study evaluated the effect of knockout serum replacement (KSR), fetal bovine serum (FBS) and bovine serum albumin (BSA) on the viability and growth of bovine secondary follicles cultured in vitro for 12 days. To this end, secondary follicles were isolated (185-202 μm) and cultured in vitro in TCM-199+ medium supplemented with KSR (5% and 10%), FBS (5% and 10%) or BSA (3 mg/ml) at 38.5°C with 5% CO2 in air. Follicular diameters were evaluated on days 0, 4, 8 and 12. After 12 days of culture, follicular survival analysis was performing by using calcein-AM and ethidium homodimer. Before and after culture, follicles were fixed in paraformaldehyde for histological evaluation. Follicular diameter at different days of culture were compared using the Kruskal-Wallis test, while the percentages of viable follicles were analyzed by chi-squared test (P < 0.05). Results showed that follicles cultured in the presence of KSR at both concentrations presented higher follicular survival rates than those cultured in control medium alone or supplemented with FBS or BSA. Conversely, the presence of KSR, BSA or FBS did not increase follicular diameter after 12 days of culture. Histology analysis showed that, among the tested treatments, follicles cultured in the presence of KSR had preserved rounded oocytes, juxtaposed granulosa cells and intact basal membrane. In conclusion, supplementation of culture medium with KSR increases the follicular survival of bovine secondary follicles cultured in vitro.</abstract><cop>England</cop><pub>Cambridge University Press</pub><pmid>31601279</pmid><doi>10.1017/S0967199419000583</doi><tpages>5</tpages><orcidid>https://orcid.org/0000-0002-5970-6177</orcidid></addata></record> |
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subjects | Bovine serum albumin Calcein Carbon dioxide Cell culture Chi-square test Embryos Fetal calf serum Follicles Gametocytes Granulosa cells Histology Laboratories Morphology Oocytes Protein sources Proteins Serum albumin Statistical tests Supplements Survival Variance analysis |
title | Supplementation of culture medium with knockout serum replacement improves the survival of bovine secondary follicles when compared with other protein sources during in vitro culture |
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