Mesenchymal stem cell isolation and characterization from human spinal ligaments
► Mesenchymal stem cells (MSCs) were isolated from human spinal ligaments. ► MSCs localized in the collagenous matrix of the spinal ligament. ► MSCs seems to be adjacent to small blood vessels. ► Analysis of MSCs may lead to elucidating the hypertrophy or ossification of them. Mesenchymal stem cells...
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| Veröffentlicht in: | Biochemical and biophysical research communications 2012-01, Vol.417 (4), p.1193-1199 |
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| creator | Asari, Toru Furukawa, Ken-Ichi Tanaka, Sunao Kudo, Hitoshi Mizukami, Hiroki Ono, Atsushi Numasawa, Takuya Kumagai, Gentaro Motomura, Shigeru Yagihashi, Soroku Toh, Satoshi |
| description | ► Mesenchymal stem cells (MSCs) were isolated from human spinal ligaments. ► MSCs localized in the collagenous matrix of the spinal ligament. ► MSCs seems to be adjacent to small blood vessels. ► Analysis of MSCs may lead to elucidating the hypertrophy or ossification of them.
Mesenchymal stem cells (MSCs) have a fibroblast-like morphology, multilineage potential, long-term viability and capacity for self-renewal. While several articles describe isolating MSCs from various human tissues, there are no reports of isolating MSCs from human spinal ligaments, and their localization in situ. If MSCs are found in human spinal ligaments, they could be used to investigate hypertrophy or ossification of spinal ligaments. To isolate and characterize MSCs from human spinal ligaments, spinal ligaments were harvested aseptically from eight patients during surgery for lumbar spinal canal stenosis and ossification of the posterior longitudinal ligament. After collagenase digestion, nucleated cells were seeded at an appropriate density to avoid colony-to-colony contact. Cells were cultured in osteogenic, adipogenic or chondrogenic media to evaluate their multilineage differentiation potential. Immunophenotypic analysis of cell surface markers was performed by flow cytometry. Spinal ligaments were processed for immunostaining using MSC-related antibodies. Cells from human spinal ligaments could be extensively expanded with limited senescence. They were able to differentiate into osteogenic, adipogenic or chondrogenic cells. Flow cytometry revealed that their phenotypic characteristics met the minimum criteria of MSCs. Immunohistochemistry revealed the localization of CD90-positive cells in the collagenous matrix of the ligament, and in adjacent small blood vessels. We isolated and expanded MSCs from human spinal ligaments and demonstrated localization of MSCs in spinal ligaments. These cells may play an indispensable role in elucidating the pathogenesis of numerous spinal diseases. |
| doi_str_mv | 10.1016/j.bbrc.2011.12.106 |
| format | Article |
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Mesenchymal stem cells (MSCs) have a fibroblast-like morphology, multilineage potential, long-term viability and capacity for self-renewal. While several articles describe isolating MSCs from various human tissues, there are no reports of isolating MSCs from human spinal ligaments, and their localization in situ. If MSCs are found in human spinal ligaments, they could be used to investigate hypertrophy or ossification of spinal ligaments. To isolate and characterize MSCs from human spinal ligaments, spinal ligaments were harvested aseptically from eight patients during surgery for lumbar spinal canal stenosis and ossification of the posterior longitudinal ligament. After collagenase digestion, nucleated cells were seeded at an appropriate density to avoid colony-to-colony contact. Cells were cultured in osteogenic, adipogenic or chondrogenic media to evaluate their multilineage differentiation potential. Immunophenotypic analysis of cell surface markers was performed by flow cytometry. Spinal ligaments were processed for immunostaining using MSC-related antibodies. Cells from human spinal ligaments could be extensively expanded with limited senescence. They were able to differentiate into osteogenic, adipogenic or chondrogenic cells. Flow cytometry revealed that their phenotypic characteristics met the minimum criteria of MSCs. Immunohistochemistry revealed the localization of CD90-positive cells in the collagenous matrix of the ligament, and in adjacent small blood vessels. We isolated and expanded MSCs from human spinal ligaments and demonstrated localization of MSCs in spinal ligaments. These cells may play an indispensable role in elucidating the pathogenesis of numerous spinal diseases.</description><identifier>ISSN: 0006-291X</identifier><identifier>EISSN: 1090-2104</identifier><identifier>DOI: 10.1016/j.bbrc.2011.12.106</identifier><identifier>PMID: 22234304</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Adipocytes - cytology ; Adipogenesis ; Cell Count ; Cell Culture Techniques ; Cell Differentiation ; Cell Separation ; Cellular Senescence ; Chondrocytes - cytology ; Differentiation ; Ectopic ossification ; Humans ; Immunohistochemistry ; Ligaments - cytology ; Localization ; Mesenchymal stem cells ; Mesenchymal Stromal Cells - cytology ; Mesenchymal Stromal Cells - physiology ; Osteoblasts - cytology ; Pathogenesis ; Spinal ligaments ; Spine</subject><ispartof>Biochemical and biophysical research communications, 2012-01, Vol.417 (4), p.1193-1199</ispartof><rights>2011 Elsevier Inc.</rights><rights>Copyright © 2011 Elsevier Inc. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c355t-56ff4681aa1cc75f6a6a1a51862902acd873937f7069d263c1bcd73dff06e3433</citedby><cites>FETCH-LOGICAL-c355t-56ff4681aa1cc75f6a6a1a51862902acd873937f7069d263c1bcd73dff06e3433</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.bbrc.2011.12.106$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22234304$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Asari, Toru</creatorcontrib><creatorcontrib>Furukawa, Ken-Ichi</creatorcontrib><creatorcontrib>Tanaka, Sunao</creatorcontrib><creatorcontrib>Kudo, Hitoshi</creatorcontrib><creatorcontrib>Mizukami, Hiroki</creatorcontrib><creatorcontrib>Ono, Atsushi</creatorcontrib><creatorcontrib>Numasawa, Takuya</creatorcontrib><creatorcontrib>Kumagai, Gentaro</creatorcontrib><creatorcontrib>Motomura, Shigeru</creatorcontrib><creatorcontrib>Yagihashi, Soroku</creatorcontrib><creatorcontrib>Toh, Satoshi</creatorcontrib><title>Mesenchymal stem cell isolation and characterization from human spinal ligaments</title><title>Biochemical and biophysical research communications</title><addtitle>Biochem Biophys Res Commun</addtitle><description>► Mesenchymal stem cells (MSCs) were isolated from human spinal ligaments. ► MSCs localized in the collagenous matrix of the spinal ligament. ► MSCs seems to be adjacent to small blood vessels. ► Analysis of MSCs may lead to elucidating the hypertrophy or ossification of them.
Mesenchymal stem cells (MSCs) have a fibroblast-like morphology, multilineage potential, long-term viability and capacity for self-renewal. While several articles describe isolating MSCs from various human tissues, there are no reports of isolating MSCs from human spinal ligaments, and their localization in situ. If MSCs are found in human spinal ligaments, they could be used to investigate hypertrophy or ossification of spinal ligaments. To isolate and characterize MSCs from human spinal ligaments, spinal ligaments were harvested aseptically from eight patients during surgery for lumbar spinal canal stenosis and ossification of the posterior longitudinal ligament. After collagenase digestion, nucleated cells were seeded at an appropriate density to avoid colony-to-colony contact. Cells were cultured in osteogenic, adipogenic or chondrogenic media to evaluate their multilineage differentiation potential. Immunophenotypic analysis of cell surface markers was performed by flow cytometry. Spinal ligaments were processed for immunostaining using MSC-related antibodies. Cells from human spinal ligaments could be extensively expanded with limited senescence. They were able to differentiate into osteogenic, adipogenic or chondrogenic cells. Flow cytometry revealed that their phenotypic characteristics met the minimum criteria of MSCs. Immunohistochemistry revealed the localization of CD90-positive cells in the collagenous matrix of the ligament, and in adjacent small blood vessels. We isolated and expanded MSCs from human spinal ligaments and demonstrated localization of MSCs in spinal ligaments. These cells may play an indispensable role in elucidating the pathogenesis of numerous spinal diseases.</description><subject>Adipocytes - cytology</subject><subject>Adipogenesis</subject><subject>Cell Count</subject><subject>Cell Culture Techniques</subject><subject>Cell Differentiation</subject><subject>Cell Separation</subject><subject>Cellular Senescence</subject><subject>Chondrocytes - cytology</subject><subject>Differentiation</subject><subject>Ectopic ossification</subject><subject>Humans</subject><subject>Immunohistochemistry</subject><subject>Ligaments - cytology</subject><subject>Localization</subject><subject>Mesenchymal stem cells</subject><subject>Mesenchymal Stromal Cells - cytology</subject><subject>Mesenchymal Stromal Cells - physiology</subject><subject>Osteoblasts - cytology</subject><subject>Pathogenesis</subject><subject>Spinal ligaments</subject><subject>Spine</subject><issn>0006-291X</issn><issn>1090-2104</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kM1KxDAYRYMozjj6Ai6kO1et-ZI2bcCNDP7BiC4U3IU0TZwM_RmTVhif3pSOLl0FLudevhyEzgEngIFdbZKydCohGCABEjJ2gOaAOY4J4PQQzTHGLCYc3mfoxPsNDmDK-DGaEUJoSnE6Ry9P2utWrXeNrCPf6yZSuq4j67ta9rZrI9lWkVpLJ1Wvnf2eQuO6JloPjWwjv7VtqNb2Qza67f0pOjKy9vps_y7Q293t6_IhXj3fPy5vVrGiWdbHGTMmZQVICUrlmWGSSZAZFIxwTKSqipxympscM14RRhWUqsppZQxmOtxOF-hy2t267nPQvheN9ePtstXd4AWHglPgMJJkIpXrvHfaiK2zjXQ7AViMIsVGjCLFKFIACRkLpYv9_FA2uvqr_JoLwPUE6PDJL6ud8MoGk7qyTqteVJ39b_8HroOEwA</recordid><startdate>20120127</startdate><enddate>20120127</enddate><creator>Asari, Toru</creator><creator>Furukawa, Ken-Ichi</creator><creator>Tanaka, Sunao</creator><creator>Kudo, Hitoshi</creator><creator>Mizukami, Hiroki</creator><creator>Ono, Atsushi</creator><creator>Numasawa, Takuya</creator><creator>Kumagai, Gentaro</creator><creator>Motomura, Shigeru</creator><creator>Yagihashi, Soroku</creator><creator>Toh, Satoshi</creator><general>Elsevier 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>7X8</scope></search><sort><creationdate>20120127</creationdate><title>Mesenchymal stem cell isolation and characterization from human spinal ligaments</title><author>Asari, Toru ; Furukawa, Ken-Ichi ; Tanaka, Sunao ; Kudo, Hitoshi ; Mizukami, Hiroki ; Ono, Atsushi ; Numasawa, Takuya ; Kumagai, Gentaro ; Motomura, Shigeru ; Yagihashi, Soroku ; Toh, Satoshi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c355t-56ff4681aa1cc75f6a6a1a51862902acd873937f7069d263c1bcd73dff06e3433</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>Adipocytes - cytology</topic><topic>Adipogenesis</topic><topic>Cell Count</topic><topic>Cell Culture Techniques</topic><topic>Cell Differentiation</topic><topic>Cell Separation</topic><topic>Cellular Senescence</topic><topic>Chondrocytes - cytology</topic><topic>Differentiation</topic><topic>Ectopic ossification</topic><topic>Humans</topic><topic>Immunohistochemistry</topic><topic>Ligaments - cytology</topic><topic>Localization</topic><topic>Mesenchymal stem cells</topic><topic>Mesenchymal Stromal Cells - cytology</topic><topic>Mesenchymal Stromal Cells - physiology</topic><topic>Osteoblasts - cytology</topic><topic>Pathogenesis</topic><topic>Spinal ligaments</topic><topic>Spine</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Asari, Toru</creatorcontrib><creatorcontrib>Furukawa, Ken-Ichi</creatorcontrib><creatorcontrib>Tanaka, Sunao</creatorcontrib><creatorcontrib>Kudo, Hitoshi</creatorcontrib><creatorcontrib>Mizukami, Hiroki</creatorcontrib><creatorcontrib>Ono, Atsushi</creatorcontrib><creatorcontrib>Numasawa, Takuya</creatorcontrib><creatorcontrib>Kumagai, Gentaro</creatorcontrib><creatorcontrib>Motomura, Shigeru</creatorcontrib><creatorcontrib>Yagihashi, Soroku</creatorcontrib><creatorcontrib>Toh, Satoshi</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>Biochemical and biophysical research communications</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Asari, Toru</au><au>Furukawa, Ken-Ichi</au><au>Tanaka, Sunao</au><au>Kudo, Hitoshi</au><au>Mizukami, Hiroki</au><au>Ono, Atsushi</au><au>Numasawa, Takuya</au><au>Kumagai, Gentaro</au><au>Motomura, Shigeru</au><au>Yagihashi, Soroku</au><au>Toh, Satoshi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Mesenchymal stem cell isolation and characterization from human spinal ligaments</atitle><jtitle>Biochemical and biophysical research communications</jtitle><addtitle>Biochem Biophys Res Commun</addtitle><date>2012-01-27</date><risdate>2012</risdate><volume>417</volume><issue>4</issue><spage>1193</spage><epage>1199</epage><pages>1193-1199</pages><issn>0006-291X</issn><eissn>1090-2104</eissn><abstract>► Mesenchymal stem cells (MSCs) were isolated from human spinal ligaments. ► MSCs localized in the collagenous matrix of the spinal ligament. ► MSCs seems to be adjacent to small blood vessels. ► Analysis of MSCs may lead to elucidating the hypertrophy or ossification of them.
Mesenchymal stem cells (MSCs) have a fibroblast-like morphology, multilineage potential, long-term viability and capacity for self-renewal. While several articles describe isolating MSCs from various human tissues, there are no reports of isolating MSCs from human spinal ligaments, and their localization in situ. If MSCs are found in human spinal ligaments, they could be used to investigate hypertrophy or ossification of spinal ligaments. To isolate and characterize MSCs from human spinal ligaments, spinal ligaments were harvested aseptically from eight patients during surgery for lumbar spinal canal stenosis and ossification of the posterior longitudinal ligament. After collagenase digestion, nucleated cells were seeded at an appropriate density to avoid colony-to-colony contact. Cells were cultured in osteogenic, adipogenic or chondrogenic media to evaluate their multilineage differentiation potential. Immunophenotypic analysis of cell surface markers was performed by flow cytometry. Spinal ligaments were processed for immunostaining using MSC-related antibodies. Cells from human spinal ligaments could be extensively expanded with limited senescence. They were able to differentiate into osteogenic, adipogenic or chondrogenic cells. Flow cytometry revealed that their phenotypic characteristics met the minimum criteria of MSCs. Immunohistochemistry revealed the localization of CD90-positive cells in the collagenous matrix of the ligament, and in adjacent small blood vessels. We isolated and expanded MSCs from human spinal ligaments and demonstrated localization of MSCs in spinal ligaments. These cells may play an indispensable role in elucidating the pathogenesis of numerous spinal diseases.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>22234304</pmid><doi>10.1016/j.bbrc.2011.12.106</doi><tpages>7</tpages></addata></record> |
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| ispartof | Biochemical and biophysical research communications, 2012-01, Vol.417 (4), p.1193-1199 |
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| subjects | Adipocytes - cytology Adipogenesis Cell Count Cell Culture Techniques Cell Differentiation Cell Separation Cellular Senescence Chondrocytes - cytology Differentiation Ectopic ossification Humans Immunohistochemistry Ligaments - cytology Localization Mesenchymal stem cells Mesenchymal Stromal Cells - cytology Mesenchymal Stromal Cells - physiology Osteoblasts - cytology Pathogenesis Spinal ligaments Spine |
| title | Mesenchymal stem cell isolation and characterization from human spinal ligaments |
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