Isolation of adipose-derived stem cells: a comparison among different methods
Adipose-derived stromal cells (ASCs) are usually isolated by digestion with collagenase. We have compared alternative methods to isolate ASCs in a more economically viable protocol. Nine protocols using red blood cells lysis buffer solution, trypsin, collagenase and centrifugation were compared; the...
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| Veröffentlicht in: | Biotechnology letters 2014-04, Vol.36 (4), p.693-702 |
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| creator | Markarian, Carolina Franke Frey, Gianna Zaffari Silveira, Maiele Dornelles Chem, Eduardo Mainieri Milani, Adriana Rosa Ely, Pedro Bins Horn, Ana Paula Nardi, Nance Beyer Camassola, Melissa |
| description | Adipose-derived stromal cells (ASCs) are usually isolated by digestion with collagenase. We have compared alternative methods to isolate ASCs in a more economically viable protocol. Nine protocols using red blood cells lysis buffer solution, trypsin, collagenase and centrifugation were compared; the isolation rate, cell viability, expansion rate, immunophenotype and differentiation in adipogenic and osteogenic lineages were analyzed. ASCs were isolated and successfully maintained by digestion with trypsin. Cells presented similar immunophenotypes, adipogenic differentiation and in vitro proliferation but an osteogenic differentiation capacity up to seven times higher than ASCs isolated by collagenase. This alternative protocol is thus efficient and more cost-effective than the commonly-used methods and may represent a promising protocol for obtaining ASCs for bone tissue engineering. |
| doi_str_mv | 10.1007/s10529-013-1425-x |
| format | Article |
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We have compared alternative methods to isolate ASCs in a more economically viable protocol. Nine protocols using red blood cells lysis buffer solution, trypsin, collagenase and centrifugation were compared; the isolation rate, cell viability, expansion rate, immunophenotype and differentiation in adipogenic and osteogenic lineages were analyzed. ASCs were isolated and successfully maintained by digestion with trypsin. Cells presented similar immunophenotypes, adipogenic differentiation and in vitro proliferation but an osteogenic differentiation capacity up to seven times higher than ASCs isolated by collagenase. This alternative protocol is thus efficient and more cost-effective than the commonly-used methods and may represent a promising protocol for obtaining ASCs for bone tissue engineering.</description><identifier>ISSN: 0141-5492</identifier><identifier>EISSN: 1573-6776</identifier><identifier>DOI: 10.1007/s10529-013-1425-x</identifier><identifier>PMID: 24322777</identifier><language>eng</language><publisher>Dordrecht: Springer-Verlag</publisher><subject>Adipocytes ; Adipose Tissue - cytology ; Applied Microbiology ; Biochemistry ; Biocompatibility ; Biomedical and Life Sciences ; Biomedical materials ; Biotechnology ; bone formation ; Bones ; Cell Proliferation ; Cell Separation - methods ; Cell Survival ; cell viability ; Centrifugation ; Collagen ; Comparative studies ; Differentiation ; Digestion ; erythrocytes ; Immunophenotyping ; Life Sciences ; lipogenesis ; Microbiology ; Original Research Paper ; Specimen Handling - methods ; Stem cells ; Stem Cells - physiology ; stromal cells ; tissue engineering ; Trypsin</subject><ispartof>Biotechnology letters, 2014-04, Vol.36 (4), p.693-702</ispartof><rights>Springer Science+Business Media Dordrecht 2013</rights><rights>Springer Science+Business Media Dordrecht 2014</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c565t-8ea7c1de062f745d2e0fc2f4922de138d42d52909b549bd761d9b9281a675d3f3</citedby><cites>FETCH-LOGICAL-c565t-8ea7c1de062f745d2e0fc2f4922de138d42d52909b549bd761d9b9281a675d3f3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s10529-013-1425-x$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s10529-013-1425-x$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,776,780,27901,27902,41464,42533,51294</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24322777$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Markarian, Carolina Franke</creatorcontrib><creatorcontrib>Frey, Gianna Zaffari</creatorcontrib><creatorcontrib>Silveira, Maiele Dornelles</creatorcontrib><creatorcontrib>Chem, Eduardo Mainieri</creatorcontrib><creatorcontrib>Milani, Adriana Rosa</creatorcontrib><creatorcontrib>Ely, Pedro Bins</creatorcontrib><creatorcontrib>Horn, Ana Paula</creatorcontrib><creatorcontrib>Nardi, Nance Beyer</creatorcontrib><creatorcontrib>Camassola, Melissa</creatorcontrib><title>Isolation of adipose-derived stem cells: a comparison among different methods</title><title>Biotechnology letters</title><addtitle>Biotechnol Lett</addtitle><addtitle>Biotechnol Lett</addtitle><description>Adipose-derived stromal cells (ASCs) are usually isolated by digestion with collagenase. We have compared alternative methods to isolate ASCs in a more economically viable protocol. Nine protocols using red blood cells lysis buffer solution, trypsin, collagenase and centrifugation were compared; the isolation rate, cell viability, expansion rate, immunophenotype and differentiation in adipogenic and osteogenic lineages were analyzed. ASCs were isolated and successfully maintained by digestion with trypsin. Cells presented similar immunophenotypes, adipogenic differentiation and in vitro proliferation but an osteogenic differentiation capacity up to seven times higher than ASCs isolated by collagenase. This alternative protocol is thus efficient and more cost-effective than the commonly-used methods and may represent a promising protocol for obtaining ASCs for bone tissue engineering.</description><subject>Adipocytes</subject><subject>Adipose Tissue - cytology</subject><subject>Applied Microbiology</subject><subject>Biochemistry</subject><subject>Biocompatibility</subject><subject>Biomedical and Life Sciences</subject><subject>Biomedical materials</subject><subject>Biotechnology</subject><subject>bone formation</subject><subject>Bones</subject><subject>Cell Proliferation</subject><subject>Cell Separation - methods</subject><subject>Cell Survival</subject><subject>cell viability</subject><subject>Centrifugation</subject><subject>Collagen</subject><subject>Comparative studies</subject><subject>Differentiation</subject><subject>Digestion</subject><subject>erythrocytes</subject><subject>Immunophenotyping</subject><subject>Life Sciences</subject><subject>lipogenesis</subject><subject>Microbiology</subject><subject>Original Research Paper</subject><subject>Specimen Handling - 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Academic</collection><collection>Biotechnology Research Abstracts</collection><jtitle>Biotechnology letters</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Markarian, Carolina Franke</au><au>Frey, Gianna Zaffari</au><au>Silveira, Maiele Dornelles</au><au>Chem, Eduardo Mainieri</au><au>Milani, Adriana Rosa</au><au>Ely, Pedro Bins</au><au>Horn, Ana Paula</au><au>Nardi, Nance Beyer</au><au>Camassola, Melissa</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Isolation of adipose-derived stem cells: a comparison among different methods</atitle><jtitle>Biotechnology letters</jtitle><stitle>Biotechnol Lett</stitle><addtitle>Biotechnol Lett</addtitle><date>2014-04-01</date><risdate>2014</risdate><volume>36</volume><issue>4</issue><spage>693</spage><epage>702</epage><pages>693-702</pages><issn>0141-5492</issn><eissn>1573-6776</eissn><abstract>Adipose-derived stromal cells (ASCs) are usually isolated by digestion with collagenase. We have compared alternative methods to isolate ASCs in a more economically viable protocol. Nine protocols using red blood cells lysis buffer solution, trypsin, collagenase and centrifugation were compared; the isolation rate, cell viability, expansion rate, immunophenotype and differentiation in adipogenic and osteogenic lineages were analyzed. ASCs were isolated and successfully maintained by digestion with trypsin. Cells presented similar immunophenotypes, adipogenic differentiation and in vitro proliferation but an osteogenic differentiation capacity up to seven times higher than ASCs isolated by collagenase. This alternative protocol is thus efficient and more cost-effective than the commonly-used methods and may represent a promising protocol for obtaining ASCs for bone tissue engineering.</abstract><cop>Dordrecht</cop><pub>Springer-Verlag</pub><pmid>24322777</pmid><doi>10.1007/s10529-013-1425-x</doi><tpages>10</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0141-5492 |
| ispartof | Biotechnology letters, 2014-04, Vol.36 (4), p.693-702 |
| issn | 0141-5492 1573-6776 |
| language | eng |
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| source | MEDLINE; Springer Nature - Complete Springer Journals |
| subjects | Adipocytes Adipose Tissue - cytology Applied Microbiology Biochemistry Biocompatibility Biomedical and Life Sciences Biomedical materials Biotechnology bone formation Bones Cell Proliferation Cell Separation - methods Cell Survival cell viability Centrifugation Collagen Comparative studies Differentiation Digestion erythrocytes Immunophenotyping Life Sciences lipogenesis Microbiology Original Research Paper Specimen Handling - methods Stem cells Stem Cells - physiology stromal cells tissue engineering Trypsin |
| title | Isolation of adipose-derived stem cells: a comparison among different methods |
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