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Two modified density gradient centrifugation methods facilitate the isolation of mouse Leydig cells
Preparation of sufficient mouse Leydig cells (LCs) with high purity is a prerequisite for investigations of the biological/pathological functions of LCs in mouse models. Density gradient centrifugation based on discontinuous Percoll gradients is an effective method (defined as regular method) for LC...
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Published in: | Preparative biochemistry & biotechnology 2023-01, Vol.53 (1), p.76-80 |
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description | Preparation of sufficient mouse Leydig cells (LCs) with high purity is a prerequisite for investigations of the biological/pathological functions of LCs in mouse models. Density gradient centrifugation based on discontinuous Percoll gradients is an effective method (defined as regular method) for LC isolation. In this study, we developed two modified methods for LC isolation and compared their performance with that of the regular method. Modified method 1 integrated the crude LCs into the 50% Percoll solution before centrifugation. Modified method 2 sequentially used 50 and 60% Percoll solutions to isolate LCs. The purity of LCs was approximately 88.4, 91.3, and 79.7% derived from the regular, modified 1, and modified 2 methods, respectively. The yields of LCs in the same respective order were approximately 1.7 × 10
5
, 3.9 × 10
5
, and 11.9 × 10
5
cells per 10
8
interstitial cells input. Modified method 1 attained higher purity and yields than those of the regular method. Although the purity of LCs was relatively low for modified method 2, it could be used before further purification by, for example, fluorescence-activated or magnetic-activated cell sorting, owing to its simplicity and high yields. Therefore, our study provided alternative methods to facilitate LC isolation in mice. |
doi_str_mv | 10.1080/10826068.2022.2039942 |
format | article |
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5
, 3.9 × 10
5
, and 11.9 × 10
5
cells per 10
8
interstitial cells input. Modified method 1 attained higher purity and yields than those of the regular method. Although the purity of LCs was relatively low for modified method 2, it could be used before further purification by, for example, fluorescence-activated or magnetic-activated cell sorting, owing to its simplicity and high yields. Therefore, our study provided alternative methods to facilitate LC isolation in mice.</description><identifier>ISSN: 1082-6068</identifier><identifier>EISSN: 1532-2297</identifier><identifier>DOI: 10.1080/10826068.2022.2039942</identifier><identifier>PMID: 35196461</identifier><language>eng</language><publisher>England: Taylor & Francis</publisher><subject>Animal models ; Animals ; Cell Separation - methods ; Centrifugation ; Centrifugation, Density Gradient - methods ; Density ; Density gradient centrifugation ; Density gradients ; Fluorescence ; Interstitial cells ; Leydig cell isolation ; Leydig Cells ; Male ; Methods ; Mice ; Percoll gradient ; Purity</subject><ispartof>Preparative biochemistry & biotechnology, 2023-01, Vol.53 (1), p.76-80</ispartof><rights>2022 Taylor & Francis Group, LLC 2022</rights><rights>2022 Taylor & Francis Group, LLC</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c394t-b8d8434c1269e7816888ff50c091b3f2534e2d85c848605d99c29380585e91113</citedby><cites>FETCH-LOGICAL-c394t-b8d8434c1269e7816888ff50c091b3f2534e2d85c848605d99c29380585e91113</cites></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/35196461$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Jiang, Jiayang</creatorcontrib><creatorcontrib>Zhou, Xiaoman</creatorcontrib><creatorcontrib>Gao, Chunliu</creatorcontrib><creatorcontrib>Ke, Rongqin</creatorcontrib><creatorcontrib>Guo, Qiwei</creatorcontrib><title>Two modified density gradient centrifugation methods facilitate the isolation of mouse Leydig cells</title><title>Preparative biochemistry & biotechnology</title><addtitle>Prep Biochem Biotechnol</addtitle><description>Preparation of sufficient mouse Leydig cells (LCs) with high purity is a prerequisite for investigations of the biological/pathological functions of LCs in mouse models. Density gradient centrifugation based on discontinuous Percoll gradients is an effective method (defined as regular method) for LC isolation. In this study, we developed two modified methods for LC isolation and compared their performance with that of the regular method. Modified method 1 integrated the crude LCs into the 50% Percoll solution before centrifugation. Modified method 2 sequentially used 50 and 60% Percoll solutions to isolate LCs. The purity of LCs was approximately 88.4, 91.3, and 79.7% derived from the regular, modified 1, and modified 2 methods, respectively. The yields of LCs in the same respective order were approximately 1.7 × 10
5
, 3.9 × 10
5
, and 11.9 × 10
5
cells per 10
8
interstitial cells input. Modified method 1 attained higher purity and yields than those of the regular method. Although the purity of LCs was relatively low for modified method 2, it could be used before further purification by, for example, fluorescence-activated or magnetic-activated cell sorting, owing to its simplicity and high yields. Therefore, our study provided alternative methods to facilitate LC isolation in mice.</description><subject>Animal models</subject><subject>Animals</subject><subject>Cell Separation - methods</subject><subject>Centrifugation</subject><subject>Centrifugation, Density Gradient - methods</subject><subject>Density</subject><subject>Density gradient centrifugation</subject><subject>Density gradients</subject><subject>Fluorescence</subject><subject>Interstitial cells</subject><subject>Leydig cell isolation</subject><subject>Leydig Cells</subject><subject>Male</subject><subject>Methods</subject><subject>Mice</subject><subject>Percoll gradient</subject><subject>Purity</subject><issn>1082-6068</issn><issn>1532-2297</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2023</creationdate><recordtype>article</recordtype><recordid>eNp9kTtPBCEAhInR-P4JGhIbm1UeCwud5uIrucRGa8LxODG7iwIbc_9eLndaWNgMFN8MEwaAM4yuMBLougrhiIsrggipQqVsyQ44xIyShhDZ7dZ7ZZo1dACOcn5HCMsOi31wQBmWvOX4EJiXrwiHaIMPzkLrxhzKCi6TtsGNBZoqKfhpqUuIIxxceYs2Q69N6EPRxcHy5mDIsd8A0dewKTs4dysbltXf9_kE7HndZ3e6PY_B6_3dy-yxmT8_PM1u542hsi3NQljR0tZgwqXrBOZCCO8ZMkjiBfWE0dYRK5gRreCIWSkNkVQgJpiTGGN6DC43uR8pfk4uFzWEvG6gR1dLKcIpERi3Ulb04g_6Hqc01naKdBwhQhHtKsU2lEkx5-S8-khh0GmlMFLrFdTPCmq9gtquUH3n2_RpMTj76_r59grcbIAw-pgG_RVTb1XRqz4mn_RoQlb0_ze-ASXplUY</recordid><startdate>20230103</startdate><enddate>20230103</enddate><creator>Jiang, Jiayang</creator><creator>Zhou, Xiaoman</creator><creator>Gao, Chunliu</creator><creator>Ke, Rongqin</creator><creator>Guo, Qiwei</creator><general>Taylor & Francis</general><general>Taylor & Francis Ltd</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>7QL</scope><scope>7QO</scope><scope>7QR</scope><scope>7T7</scope><scope>7TK</scope><scope>7TM</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>M7N</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20230103</creationdate><title>Two modified density gradient centrifugation methods facilitate the isolation of mouse Leydig cells</title><author>Jiang, Jiayang ; Zhou, Xiaoman ; Gao, Chunliu ; Ke, Rongqin ; Guo, Qiwei</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c394t-b8d8434c1269e7816888ff50c091b3f2534e2d85c848605d99c29380585e91113</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2023</creationdate><topic>Animal models</topic><topic>Animals</topic><topic>Cell Separation - methods</topic><topic>Centrifugation</topic><topic>Centrifugation, Density Gradient - methods</topic><topic>Density</topic><topic>Density gradient centrifugation</topic><topic>Density gradients</topic><topic>Fluorescence</topic><topic>Interstitial cells</topic><topic>Leydig cell isolation</topic><topic>Leydig Cells</topic><topic>Male</topic><topic>Methods</topic><topic>Mice</topic><topic>Percoll gradient</topic><topic>Purity</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Jiang, Jiayang</creatorcontrib><creatorcontrib>Zhou, Xiaoman</creatorcontrib><creatorcontrib>Gao, Chunliu</creatorcontrib><creatorcontrib>Ke, Rongqin</creatorcontrib><creatorcontrib>Guo, Qiwei</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Preparative biochemistry & biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Jiang, Jiayang</au><au>Zhou, Xiaoman</au><au>Gao, Chunliu</au><au>Ke, Rongqin</au><au>Guo, Qiwei</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Two modified density gradient centrifugation methods facilitate the isolation of mouse Leydig cells</atitle><jtitle>Preparative biochemistry & biotechnology</jtitle><addtitle>Prep Biochem Biotechnol</addtitle><date>2023-01-03</date><risdate>2023</risdate><volume>53</volume><issue>1</issue><spage>76</spage><epage>80</epage><pages>76-80</pages><issn>1082-6068</issn><eissn>1532-2297</eissn><notes>ObjectType-Article-1</notes><notes>SourceType-Scholarly Journals-1</notes><notes>ObjectType-Feature-2</notes><notes>content type line 23</notes><abstract>Preparation of sufficient mouse Leydig cells (LCs) with high purity is a prerequisite for investigations of the biological/pathological functions of LCs in mouse models. Density gradient centrifugation based on discontinuous Percoll gradients is an effective method (defined as regular method) for LC isolation. In this study, we developed two modified methods for LC isolation and compared their performance with that of the regular method. Modified method 1 integrated the crude LCs into the 50% Percoll solution before centrifugation. Modified method 2 sequentially used 50 and 60% Percoll solutions to isolate LCs. The purity of LCs was approximately 88.4, 91.3, and 79.7% derived from the regular, modified 1, and modified 2 methods, respectively. The yields of LCs in the same respective order were approximately 1.7 × 10
5
, 3.9 × 10
5
, and 11.9 × 10
5
cells per 10
8
interstitial cells input. Modified method 1 attained higher purity and yields than those of the regular method. Although the purity of LCs was relatively low for modified method 2, it could be used before further purification by, for example, fluorescence-activated or magnetic-activated cell sorting, owing to its simplicity and high yields. Therefore, our study provided alternative methods to facilitate LC isolation in mice.</abstract><cop>England</cop><pub>Taylor & Francis</pub><pmid>35196461</pmid><doi>10.1080/10826068.2022.2039942</doi><tpages>5</tpages></addata></record> |
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subjects | Animal models Animals Cell Separation - methods Centrifugation Centrifugation, Density Gradient - methods Density Density gradient centrifugation Density gradients Fluorescence Interstitial cells Leydig cell isolation Leydig Cells Male Methods Mice Percoll gradient Purity |
title | Two modified density gradient centrifugation methods facilitate the isolation of mouse Leydig cells |
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