Short exposure to collagenase and coculture with mouse embryonic pancreas improve human dermal fibroblast culture
The dermal fibroblast as a major component of connective tissue has attracted much attention in the past few years, and application of these very fast growing cells in several fields has been intensively studied. Isolating dermal fibroblasts is an appropriate way to expand these fast growing cells i...
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| Veröffentlicht in: | Biotechnology and applied biochemistry 2012-05, Vol.59 (3), p.254-261 |
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| creator | Pandamooz, Sareh Hadipour, Abbas Akhavan-Niaki, Haleh Pourghasem, Mohsen Abedian, Zeinab Ardekani, Ali Motevallizadeh Golpour, Monireh Hassan, Zuhair Mohammad Mostafazadeh, Amrollah |
| description | The dermal fibroblast as a major component of connective tissue has attracted much attention in the past few years, and application of these very fast growing cells in several fields has been intensively studied. Isolating dermal fibroblasts is an appropriate way to expand these fast growing cells in vitro. Although using a dissociated fibroblast culture method is more convenient than skin explant culture, its enzymatic digestion is critical because a large number of cells can be lost over prolonged exposure to collagenase. This study was performed to increase the number of viable cells after digestion of fresh human foreskin of donors aged from 1 to 3 months with collagenase and also by to design a coculture system for resuscitation of the injured fibroblast. Our results demonstrate that we can maximize cell yield while maintaining cell viability by cutting the specimens into very small pieces (1–2 mm3) after removing the epidermal layer with dispase II and also by collecting released cells every 20 Min subsequent to digesting the dermal layer with collagenase. Moreover, our data strongly indicate that coculturing of isolated fibroblasts with embryonic pancreas explants can enhance the rate of proliferation in cultured fibroblasts. |
| doi_str_mv | 10.1002/bab.1020 |
| format | Article |
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Isolating dermal fibroblasts is an appropriate way to expand these fast growing cells in vitro. Although using a dissociated fibroblast culture method is more convenient than skin explant culture, its enzymatic digestion is critical because a large number of cells can be lost over prolonged exposure to collagenase. This study was performed to increase the number of viable cells after digestion of fresh human foreskin of donors aged from 1 to 3 months with collagenase and also by to design a coculture system for resuscitation of the injured fibroblast. Our results demonstrate that we can maximize cell yield while maintaining cell viability by cutting the specimens into very small pieces (1–2 mm3) after removing the epidermal layer with dispase II and also by collecting released cells every 20 Min subsequent to digesting the dermal layer with collagenase. Moreover, our data strongly indicate that coculturing of isolated fibroblasts with embryonic pancreas explants can enhance the rate of proliferation in cultured fibroblasts.</description><identifier>ISSN: 0885-4513</identifier><identifier>EISSN: 1470-8744</identifier><identifier>DOI: 10.1002/bab.1020</identifier><identifier>PMID: 23586836</identifier><identifier>CODEN: BABIEC</identifier><language>eng</language><publisher>Hoboken: Wiley Subscription Services, Inc., A Wiley Company</publisher><subject>Animals ; Biological and medical sciences ; Biotechnology ; Cell Culture Techniques ; Cell Survival ; Coculture Techniques ; coculturing ; collagenase ; Collagenases - metabolism ; dermal fibroblasts ; Embryonic Stem Cells - cytology ; Embryonic Stem Cells - drug effects ; enzymatic digestion ; Fibroblasts - cytology ; foreskin ; Foreskin - cytology ; Fundamental and applied biological sciences. Psychology ; Humans ; Male ; Mice ; Pancreas - cytology ; pancreas explants ; Time Factors</subject><ispartof>Biotechnology and applied biochemistry, 2012-05, Vol.59 (3), p.254-261</ispartof><rights>Copyright © 2012 International Union of Biochemistry and Molecular Biology, Inc.</rights><rights>2015 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4500-fac0996ace5d08bf02485c8d6a8187c45515e668b31d592e286ce2ea3707000b3</citedby><cites>FETCH-LOGICAL-c4500-fac0996ace5d08bf02485c8d6a8187c45515e668b31d592e286ce2ea3707000b3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fbab.1020$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fbab.1020$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1417,27924,27925,45574,45575</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=26006560$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23586836$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Pandamooz, Sareh</creatorcontrib><creatorcontrib>Hadipour, Abbas</creatorcontrib><creatorcontrib>Akhavan-Niaki, Haleh</creatorcontrib><creatorcontrib>Pourghasem, Mohsen</creatorcontrib><creatorcontrib>Abedian, Zeinab</creatorcontrib><creatorcontrib>Ardekani, Ali Motevallizadeh</creatorcontrib><creatorcontrib>Golpour, Monireh</creatorcontrib><creatorcontrib>Hassan, Zuhair Mohammad</creatorcontrib><creatorcontrib>Mostafazadeh, Amrollah</creatorcontrib><title>Short exposure to collagenase and coculture with mouse embryonic pancreas improve human dermal fibroblast culture</title><title>Biotechnology and applied biochemistry</title><addtitle>Biotechnology and Applied Biochemistry</addtitle><description>The dermal fibroblast as a major component of connective tissue has attracted much attention in the past few years, and application of these very fast growing cells in several fields has been intensively studied. Isolating dermal fibroblasts is an appropriate way to expand these fast growing cells in vitro. Although using a dissociated fibroblast culture method is more convenient than skin explant culture, its enzymatic digestion is critical because a large number of cells can be lost over prolonged exposure to collagenase. This study was performed to increase the number of viable cells after digestion of fresh human foreskin of donors aged from 1 to 3 months with collagenase and also by to design a coculture system for resuscitation of the injured fibroblast. Our results demonstrate that we can maximize cell yield while maintaining cell viability by cutting the specimens into very small pieces (1–2 mm3) after removing the epidermal layer with dispase II and also by collecting released cells every 20 Min subsequent to digesting the dermal layer with collagenase. Moreover, our data strongly indicate that coculturing of isolated fibroblasts with embryonic pancreas explants can enhance the rate of proliferation in cultured fibroblasts.</description><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Cell Culture Techniques</subject><subject>Cell Survival</subject><subject>Coculture Techniques</subject><subject>coculturing</subject><subject>collagenase</subject><subject>Collagenases - metabolism</subject><subject>dermal fibroblasts</subject><subject>Embryonic Stem Cells - cytology</subject><subject>Embryonic Stem Cells - drug effects</subject><subject>enzymatic digestion</subject><subject>Fibroblasts - cytology</subject><subject>foreskin</subject><subject>Foreskin - cytology</subject><subject>Fundamental and applied biological sciences. 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| ispartof | Biotechnology and applied biochemistry, 2012-05, Vol.59 (3), p.254-261 |
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| subjects | Animals Biological and medical sciences Biotechnology Cell Culture Techniques Cell Survival Coculture Techniques coculturing collagenase Collagenases - metabolism dermal fibroblasts Embryonic Stem Cells - cytology Embryonic Stem Cells - drug effects enzymatic digestion Fibroblasts - cytology foreskin Foreskin - cytology Fundamental and applied biological sciences. Psychology Humans Male Mice Pancreas - cytology pancreas explants Time Factors |
| title | Short exposure to collagenase and coculture with mouse embryonic pancreas improve human dermal fibroblast culture |
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