Strategy towards tailored donor tissue-specific pancreatic islet isolation
Optimizing the collagenase G (ColG):collagenase H (ColH) ratio is a key strategy for achieving tailored donor-tissue specific islet isolation. Collagen V (Col V) and collagen III (Col III) are crucial target matrices of ColG and ColH, respectively. We herein investigated the relevance between the ex...
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| Veröffentlicht in: | PloS one 2019-05, Vol.14 (5), p.e0216136-e0216136 |
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| creator | Miyazaki, Yuki Murayama, Kazutaka Fathi, Ibrahim Imura, Takehiro Yamagata, Youhei Watanabe, Kimiko Maeda, Hiroshi Inagaki, Akiko Igarashi, Yasuhiro Miyagi, Shigehito Shima, Hiroki Igarashi, Kazuhiko Kamei, Takashi Unno, Michiaki Goto, Masafumi |
| description | Optimizing the collagenase G (ColG):collagenase H (ColH) ratio is a key strategy for achieving tailored donor-tissue specific islet isolation. Collagen V (Col V) and collagen III (Col III) are crucial target matrices of ColG and ColH, respectively. We herein investigated the relevance between the expression of target matrices in pancreatic tissues and influence of ColG:ColH ratio on islet isolation outcome.
Islet isolation was performed in Lewis and SD rats using different ColG:ColH ratios (5:1, 1:1 and 1:5; n = 7/group). The composition of Col III and Col V was examined using immunohistochemical staining, real-time polymerase chain reaction (PCR), Western blotting and mass spectrometry. Chain types in collagen I (Col I) were also assessed using mass spectrometry.
No beneficial effects were observed by increasing the ColG amount, irrespective of the rat strain. In contrast, the islet yield in Lewis rats was considerably increased by high amounts of ColH but decreased in SD rats, suggesting that Lewis pancreas contains more Col III than SD pancreas. Neither immunohistochemical nor real-time PCR showed correlation with isolation outcome. However, Western blotting revealed that Lewis contained considerably higher amount of Col III than SD (p = 0.10). Likewise, Col-I(α1)/Col-III(α1) and Col-I(α2)/Col-III(α1) were significantly lower in Lewis than in SD rats (p = 0.007, respectively). Furthermore, the isolation outcome was considerably correlated with the composition of homotrimeric Col I.
The Col III expression and the composition of homotrimeric Col I in pancreatic tissues determined using mass analyses appeared useful for optimizing the ColG:ColH ratio in islet isolation. |
| doi_str_mv | 10.1371/journal.pone.0216136 |
| format | Article |
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Islet isolation was performed in Lewis and SD rats using different ColG:ColH ratios (5:1, 1:1 and 1:5; n = 7/group). The composition of Col III and Col V was examined using immunohistochemical staining, real-time polymerase chain reaction (PCR), Western blotting and mass spectrometry. Chain types in collagen I (Col I) were also assessed using mass spectrometry.
No beneficial effects were observed by increasing the ColG amount, irrespective of the rat strain. In contrast, the islet yield in Lewis rats was considerably increased by high amounts of ColH but decreased in SD rats, suggesting that Lewis pancreas contains more Col III than SD pancreas. Neither immunohistochemical nor real-time PCR showed correlation with isolation outcome. However, Western blotting revealed that Lewis contained considerably higher amount of Col III than SD (p = 0.10). Likewise, Col-I(α1)/Col-III(α1) and Col-I(α2)/Col-III(α1) were significantly lower in Lewis than in SD rats (p = 0.007, respectively). Furthermore, the isolation outcome was considerably correlated with the composition of homotrimeric Col I.
The Col III expression and the composition of homotrimeric Col I in pancreatic tissues determined using mass analyses appeared useful for optimizing the ColG:ColH ratio in islet isolation.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0216136</identifier><identifier>PMID: 31075114</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Agriculture ; Animals ; Aprotinin ; Biochemistry ; Biology and Life Sciences ; Collagen ; Collagen (type I) ; Collagen (type III) ; Collagen - metabolism ; Collagenase ; Collagenases - metabolism ; Composition ; Diabetes ; EDTA ; Enzymes ; Extracellular matrix ; Health aspects ; Immunohistochemistry ; Islet cell transplantation ; Islets of Langerhans - cytology ; Islets of Langerhans - metabolism ; Islets of Langerhans Transplantation - methods ; Laboratory animals ; Mass spectrometry ; Mass spectroscopy ; Medicine ; Medicine and Health Sciences ; Molecular structure ; Pancreas ; Peptides ; Polymerase chain reaction ; Proteins ; Proteomics ; Rats ; Rats, Inbred Lew ; Rats, Sprague-Dawley ; Real time ; Research and Analysis Methods ; Safety and security measures ; Scientific imaging ; Spectroscopy ; Surgery ; Tissue Donors ; Tissues ; Transplants & implants ; University graduates ; Western blotting</subject><ispartof>PloS one, 2019-05, Vol.14 (5), p.e0216136-e0216136</ispartof><rights>COPYRIGHT 2019 Public Library of Science</rights><rights>2019 Miyazaki et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2019 Miyazaki et al 2019 Miyazaki et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c802t-eaf3cd3afd7b6238334cfdfef2687e30f46ef806db5fc83ad0197b02fd4fd8d23</citedby><cites>FETCH-LOGICAL-c802t-eaf3cd3afd7b6238334cfdfef2687e30f46ef806db5fc83ad0197b02fd4fd8d23</cites><orcidid>0000-0003-2412-1854 ; 0000-0003-0561-4135</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6510438/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6510438/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,315,729,782,786,866,887,2106,2932,23875,27933,27934,53800,53802</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/31075114$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Asakura, Atsushi</contributor><creatorcontrib>Miyazaki, Yuki</creatorcontrib><creatorcontrib>Murayama, Kazutaka</creatorcontrib><creatorcontrib>Fathi, Ibrahim</creatorcontrib><creatorcontrib>Imura, Takehiro</creatorcontrib><creatorcontrib>Yamagata, Youhei</creatorcontrib><creatorcontrib>Watanabe, Kimiko</creatorcontrib><creatorcontrib>Maeda, Hiroshi</creatorcontrib><creatorcontrib>Inagaki, Akiko</creatorcontrib><creatorcontrib>Igarashi, Yasuhiro</creatorcontrib><creatorcontrib>Miyagi, Shigehito</creatorcontrib><creatorcontrib>Shima, Hiroki</creatorcontrib><creatorcontrib>Igarashi, Kazuhiko</creatorcontrib><creatorcontrib>Kamei, Takashi</creatorcontrib><creatorcontrib>Unno, Michiaki</creatorcontrib><creatorcontrib>Goto, Masafumi</creatorcontrib><title>Strategy towards tailored donor tissue-specific pancreatic islet isolation</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Optimizing the collagenase G (ColG):collagenase H (ColH) ratio is a key strategy for achieving tailored donor-tissue specific islet isolation. Collagen V (Col V) and collagen III (Col III) are crucial target matrices of ColG and ColH, respectively. We herein investigated the relevance between the expression of target matrices in pancreatic tissues and influence of ColG:ColH ratio on islet isolation outcome.
Islet isolation was performed in Lewis and SD rats using different ColG:ColH ratios (5:1, 1:1 and 1:5; n = 7/group). The composition of Col III and Col V was examined using immunohistochemical staining, real-time polymerase chain reaction (PCR), Western blotting and mass spectrometry. Chain types in collagen I (Col I) were also assessed using mass spectrometry.
No beneficial effects were observed by increasing the ColG amount, irrespective of the rat strain. In contrast, the islet yield in Lewis rats was considerably increased by high amounts of ColH but decreased in SD rats, suggesting that Lewis pancreas contains more Col III than SD pancreas. Neither immunohistochemical nor real-time PCR showed correlation with isolation outcome. However, Western blotting revealed that Lewis contained considerably higher amount of Col III than SD (p = 0.10). Likewise, Col-I(α1)/Col-III(α1) and Col-I(α2)/Col-III(α1) were significantly lower in Lewis than in SD rats (p = 0.007, respectively). Furthermore, the isolation outcome was considerably correlated with the composition of homotrimeric Col I.
The Col III expression and the composition of homotrimeric Col I in pancreatic tissues determined using mass analyses appeared useful for optimizing the ColG:ColH ratio in islet isolation.</description><subject>Agriculture</subject><subject>Animals</subject><subject>Aprotinin</subject><subject>Biochemistry</subject><subject>Biology and Life Sciences</subject><subject>Collagen</subject><subject>Collagen (type I)</subject><subject>Collagen (type III)</subject><subject>Collagen - metabolism</subject><subject>Collagenase</subject><subject>Collagenases - metabolism</subject><subject>Composition</subject><subject>Diabetes</subject><subject>EDTA</subject><subject>Enzymes</subject><subject>Extracellular matrix</subject><subject>Health aspects</subject><subject>Immunohistochemistry</subject><subject>Islet cell transplantation</subject><subject>Islets of Langerhans - cytology</subject><subject>Islets of Langerhans - metabolism</subject><subject>Islets of Langerhans Transplantation - methods</subject><subject>Laboratory animals</subject><subject>Mass spectrometry</subject><subject>Mass spectroscopy</subject><subject>Medicine</subject><subject>Medicine and Health Sciences</subject><subject>Molecular structure</subject><subject>Pancreas</subject><subject>Peptides</subject><subject>Polymerase chain reaction</subject><subject>Proteins</subject><subject>Proteomics</subject><subject>Rats</subject><subject>Rats, Inbred Lew</subject><subject>Rats, Sprague-Dawley</subject><subject>Real time</subject><subject>Research and Analysis Methods</subject><subject>Safety and security measures</subject><subject>Scientific imaging</subject><subject>Spectroscopy</subject><subject>Surgery</subject><subject>Tissue Donors</subject><subject>Tissues</subject><subject>Transplants & implants</subject><subject>University graduates</subject><subject>Western 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towards tailored donor tissue-specific pancreatic islet isolation</title><author>Miyazaki, Yuki ; Murayama, Kazutaka ; Fathi, Ibrahim ; Imura, Takehiro ; Yamagata, Youhei ; Watanabe, Kimiko ; Maeda, Hiroshi ; Inagaki, Akiko ; Igarashi, Yasuhiro ; Miyagi, Shigehito ; Shima, Hiroki ; Igarashi, Kazuhiko ; Kamei, Takashi ; Unno, Michiaki ; Goto, Masafumi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c802t-eaf3cd3afd7b6238334cfdfef2687e30f46ef806db5fc83ad0197b02fd4fd8d23</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2019</creationdate><topic>Agriculture</topic><topic>Animals</topic><topic>Aprotinin</topic><topic>Biochemistry</topic><topic>Biology and Life Sciences</topic><topic>Collagen</topic><topic>Collagen (type I)</topic><topic>Collagen (type III)</topic><topic>Collagen - metabolism</topic><topic>Collagenase</topic><topic>Collagenases - 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Shigehito</au><au>Shima, Hiroki</au><au>Igarashi, Kazuhiko</au><au>Kamei, Takashi</au><au>Unno, Michiaki</au><au>Goto, Masafumi</au><au>Asakura, Atsushi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Strategy towards tailored donor tissue-specific pancreatic islet isolation</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2019-05-10</date><risdate>2019</risdate><volume>14</volume><issue>5</issue><spage>e0216136</spage><epage>e0216136</epage><pages>e0216136-e0216136</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Optimizing the collagenase G (ColG):collagenase H (ColH) ratio is a key strategy for achieving tailored donor-tissue specific islet isolation. Collagen V (Col V) and collagen III (Col III) are crucial target matrices of ColG and ColH, respectively. We herein investigated the relevance between the expression of target matrices in pancreatic tissues and influence of ColG:ColH ratio on islet isolation outcome.
Islet isolation was performed in Lewis and SD rats using different ColG:ColH ratios (5:1, 1:1 and 1:5; n = 7/group). The composition of Col III and Col V was examined using immunohistochemical staining, real-time polymerase chain reaction (PCR), Western blotting and mass spectrometry. Chain types in collagen I (Col I) were also assessed using mass spectrometry.
No beneficial effects were observed by increasing the ColG amount, irrespective of the rat strain. In contrast, the islet yield in Lewis rats was considerably increased by high amounts of ColH but decreased in SD rats, suggesting that Lewis pancreas contains more Col III than SD pancreas. Neither immunohistochemical nor real-time PCR showed correlation with isolation outcome. However, Western blotting revealed that Lewis contained considerably higher amount of Col III than SD (p = 0.10). Likewise, Col-I(α1)/Col-III(α1) and Col-I(α2)/Col-III(α1) were significantly lower in Lewis than in SD rats (p = 0.007, respectively). Furthermore, the isolation outcome was considerably correlated with the composition of homotrimeric Col I.
The Col III expression and the composition of homotrimeric Col I in pancreatic tissues determined using mass analyses appeared useful for optimizing the ColG:ColH ratio in islet isolation.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>31075114</pmid><doi>10.1371/journal.pone.0216136</doi><tpages>e0216136</tpages><orcidid>https://orcid.org/0000-0003-2412-1854</orcidid><orcidid>https://orcid.org/0000-0003-0561-4135</orcidid><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2019-05, Vol.14 (5), p.e0216136-e0216136 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_2223020570 |
| source | Public Library of Science (PLoS) Journals Open Access; MEDLINE; DOAJ Directory of Open Access Journals; EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Agriculture Animals Aprotinin Biochemistry Biology and Life Sciences Collagen Collagen (type I) Collagen (type III) Collagen - metabolism Collagenase Collagenases - metabolism Composition Diabetes EDTA Enzymes Extracellular matrix Health aspects Immunohistochemistry Islet cell transplantation Islets of Langerhans - cytology Islets of Langerhans - metabolism Islets of Langerhans Transplantation - methods Laboratory animals Mass spectrometry Mass spectroscopy Medicine Medicine and Health Sciences Molecular structure Pancreas Peptides Polymerase chain reaction Proteins Proteomics Rats Rats, Inbred Lew Rats, Sprague-Dawley Real time Research and Analysis Methods Safety and security measures Scientific imaging Spectroscopy Surgery Tissue Donors Tissues Transplants & implants University graduates Western blotting |
| title | Strategy towards tailored donor tissue-specific pancreatic islet isolation |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-11-29T20%3A45%3A07IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Strategy%20towards%20tailored%20donor%20tissue-specific%20pancreatic%20islet%20isolation&rft.jtitle=PloS%20one&rft.au=Miyazaki,%20Yuki&rft.date=2019-05-10&rft.volume=14&rft.issue=5&rft.spage=e0216136&rft.epage=e0216136&rft.pages=e0216136-e0216136&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0216136&rft_dat=%3Cgale_plos_%3EA585044236%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2223020570&rft_id=info:pmid/31075114&rft_galeid=A585044236&rft_doaj_id=oai_doaj_org_article_b75e932c6d3c4669ad00cc70fd81c633&rfr_iscdi=true |