Quantitative Assessment of β-Cell Apoptosis and Cell Composition of Isolated, Undisrupted Human Islets by Laser Scanning Cytometry
Assays for assessing human islet cell quality, which provide results before transplantation, would be beneficial to improve the outcomes for islet transplantation therapy. Parameters such as percent β-cell apoptosis and cell composition are found to vary markedly between different islet preparations...
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| Veröffentlicht in: | Transplantation 2010-10, Vol.90 (8), p.836-842 |
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| creator | TODOROV, Ivan NAIR, Indu FERRERI, Kevin AL-ABDULLAH, Ismail H MULLEN, Yoko KANDEEL, Fouad AVAKIAN-MANSOORIAN, Alina RAWSON, Jeffrey OMORI, Keiko ITO, Taihei VALIENTE, Luis IGLESIAS-MEZA, Itzia ORR, Chris SHIANG, Keh-Dong |
| description | Assays for assessing human islet cell quality, which provide results before transplantation, would be beneficial to improve the outcomes for islet transplantation therapy. Parameters such as percent β-cell apoptosis and cell composition are found to vary markedly between different islet preparations and may serve as markers of islet quality. We have developed fluorescence-based assays using laser scanning cytometry for assessing β-cell apoptosis and islet cell composition on serial sections of intact isolated islets.
Isolated human islets were fixed in formalin and embedded in paraffin. Serial sections were immunostained for the pancreatic hormones and acinar and ductal cell markers. DNA fragmentation was used to label apoptotic cells. Stained cells were quantified using an iCys laser scanning cytometer.
Islet preparations from 102 human pancreatic islet isolations were analyzed. For the whole set of islet preparations, we found a mean islet cell composition of 54.5%±1.2% insulin-positive, 33.9%±1.2% glucagon, 12.1%±0.7% somatostatin, and 1.5%±0.2% pancreatic polypeptide-positive cells. The apoptotic β cells were 2.85%±0.4% with a range of 0.27% to 18.3%. The percentage of apoptotic β cells correlated well (P |
| doi_str_mv | 10.1097/TP.0b013e3181f1db5d |
| format | Article |
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Isolated human islets were fixed in formalin and embedded in paraffin. Serial sections were immunostained for the pancreatic hormones and acinar and ductal cell markers. DNA fragmentation was used to label apoptotic cells. Stained cells were quantified using an iCys laser scanning cytometer.
Islet preparations from 102 human pancreatic islet isolations were analyzed. For the whole set of islet preparations, we found a mean islet cell composition of 54.5%±1.2% insulin-positive, 33.9%±1.2% glucagon, 12.1%±0.7% somatostatin, and 1.5%±0.2% pancreatic polypeptide-positive cells. The apoptotic β cells were 2.85%±0.4% with a range of 0.27% to 18.3%. The percentage of apoptotic β cells correlated well (P<0.0001, n=59) with results obtained in vivo by transplantation of the corresponding islets in diabetic NODscid mice.
The analysis of whole, nondissociated islets for cell composition and β-cell apoptosis using laser scanning cytometry gives reliable and reproducible results and could be performed both before islet transplantation and on preserved cell blocks at any time in future. Thus, they can be a powerful tool for islet quality assessment.</description><identifier>ISSN: 0041-1337</identifier><identifier>EISSN: 1534-6080</identifier><identifier>DOI: 10.1097/TP.0b013e3181f1db5d</identifier><identifier>PMID: 20697327</identifier><identifier>CODEN: TRPLAU</identifier><language>eng</language><publisher>Hagerstown, MD: Lippincott Williams & Wilkins</publisher><subject>Apoptosis ; Biological and medical sciences ; Fundamental and applied biological sciences. Psychology ; Fundamental immunology ; Glucagon - metabolism ; Humans ; Insulin - metabolism ; Insulin Secretion ; Insulin-Secreting Cells - metabolism ; Islets of Langerhans - cytology ; Islets of Langerhans - physiology ; Medical sciences ; Pancreatic Polypeptide - metabolism ; Somatostatin - metabolism ; Surgery (general aspects). Transplantations, organ and tissue grafts. Graft diseases ; Tissue, organ and graft immunology</subject><ispartof>Transplantation, 2010-10, Vol.90 (8), p.836-842</ispartof><rights>2015 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c434t-e4a780d984f2f18fff753151fd5741bd122c730fcfa1a4a53c0376b526fb4b1a3</citedby><cites>FETCH-LOGICAL-c434t-e4a780d984f2f18fff753151fd5741bd122c730fcfa1a4a53c0376b526fb4b1a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,780,784,885,27923,27924</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=23378247$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20697327$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>TODOROV, Ivan</creatorcontrib><creatorcontrib>NAIR, Indu</creatorcontrib><creatorcontrib>FERRERI, Kevin</creatorcontrib><creatorcontrib>AL-ABDULLAH, Ismail H</creatorcontrib><creatorcontrib>MULLEN, Yoko</creatorcontrib><creatorcontrib>KANDEEL, Fouad</creatorcontrib><creatorcontrib>AVAKIAN-MANSOORIAN, Alina</creatorcontrib><creatorcontrib>RAWSON, Jeffrey</creatorcontrib><creatorcontrib>OMORI, Keiko</creatorcontrib><creatorcontrib>ITO, Taihei</creatorcontrib><creatorcontrib>VALIENTE, Luis</creatorcontrib><creatorcontrib>IGLESIAS-MEZA, Itzia</creatorcontrib><creatorcontrib>ORR, Chris</creatorcontrib><creatorcontrib>SHIANG, Keh-Dong</creatorcontrib><title>Quantitative Assessment of β-Cell Apoptosis and Cell Composition of Isolated, Undisrupted Human Islets by Laser Scanning Cytometry</title><title>Transplantation</title><addtitle>Transplantation</addtitle><description>Assays for assessing human islet cell quality, which provide results before transplantation, would be beneficial to improve the outcomes for islet transplantation therapy. Parameters such as percent β-cell apoptosis and cell composition are found to vary markedly between different islet preparations and may serve as markers of islet quality. We have developed fluorescence-based assays using laser scanning cytometry for assessing β-cell apoptosis and islet cell composition on serial sections of intact isolated islets.
Isolated human islets were fixed in formalin and embedded in paraffin. Serial sections were immunostained for the pancreatic hormones and acinar and ductal cell markers. DNA fragmentation was used to label apoptotic cells. Stained cells were quantified using an iCys laser scanning cytometer.
Islet preparations from 102 human pancreatic islet isolations were analyzed. For the whole set of islet preparations, we found a mean islet cell composition of 54.5%±1.2% insulin-positive, 33.9%±1.2% glucagon, 12.1%±0.7% somatostatin, and 1.5%±0.2% pancreatic polypeptide-positive cells. The apoptotic β cells were 2.85%±0.4% with a range of 0.27% to 18.3%. The percentage of apoptotic β cells correlated well (P<0.0001, n=59) with results obtained in vivo by transplantation of the corresponding islets in diabetic NODscid mice.
The analysis of whole, nondissociated islets for cell composition and β-cell apoptosis using laser scanning cytometry gives reliable and reproducible results and could be performed both before islet transplantation and on preserved cell blocks at any time in future. Thus, they can be a powerful tool for islet quality assessment.</description><subject>Apoptosis</subject><subject>Biological and medical sciences</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Fundamental immunology</subject><subject>Glucagon - metabolism</subject><subject>Humans</subject><subject>Insulin - metabolism</subject><subject>Insulin Secretion</subject><subject>Insulin-Secreting Cells - metabolism</subject><subject>Islets of Langerhans - cytology</subject><subject>Islets of Langerhans - physiology</subject><subject>Medical sciences</subject><subject>Pancreatic Polypeptide - metabolism</subject><subject>Somatostatin - metabolism</subject><subject>Surgery (general aspects). Transplantations, organ and tissue grafts. Graft diseases</subject><subject>Tissue, organ and graft immunology</subject><issn>0041-1337</issn><issn>1534-6080</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpdkd9uFCEUxonR2G31CUwMN6Y3ToUBlpkbk81EbZNNrHF7Tc4wUDEzMALTZK99Ix_EZyq1a_1zRTjf73znwIfQC0rOKGnlm93lGekJZYbRhlo69GJ4hFZUMF6tSUMeoxUhnFaUMXmEjlP6SggRTMqn6Kgm61ayWq7Q908L-OwyZHdj8CYlk9JkfMbB4p8_qs6MI97MYc4huYTBD_hXqQvTXCrZBX9HXqQwQjbDa3zlB5fiMpcLPl8m8EUbTU643-MtJBPxZw3eO3-Nu30Ok8lx_ww9sTAm8_xwnqCr9-923Xm1_fjhottsK80Zz5XhIBsytA23taWNtVYKRgW1g5Cc9gOtay0ZsdoCBQ6CacLkuhf12va8p8BO0Nt733npJzPo8swIo5qjmyDuVQCn_lW8-6Kuw42qW9FSRovB6cEghm-LSVlNLunyH-BNWJKSom3kmteskOye1DGkFI19mEKJuktP7S7V_-mVrpd_L_jQ8zuuArw6AJA0jDaC1y794UrUTc0luwWYbqhj</recordid><startdate>20101027</startdate><enddate>20101027</enddate><creator>TODOROV, Ivan</creator><creator>NAIR, Indu</creator><creator>FERRERI, Kevin</creator><creator>AL-ABDULLAH, Ismail H</creator><creator>MULLEN, Yoko</creator><creator>KANDEEL, Fouad</creator><creator>AVAKIAN-MANSOORIAN, Alina</creator><creator>RAWSON, Jeffrey</creator><creator>OMORI, Keiko</creator><creator>ITO, Taihei</creator><creator>VALIENTE, Luis</creator><creator>IGLESIAS-MEZA, Itzia</creator><creator>ORR, Chris</creator><creator>SHIANG, Keh-Dong</creator><general>Lippincott Williams & Wilkins</general><scope>IQODW</scope><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><scope>5PM</scope></search><sort><creationdate>20101027</creationdate><title>Quantitative Assessment of β-Cell Apoptosis and Cell Composition of Isolated, Undisrupted Human Islets by Laser Scanning Cytometry</title><author>TODOROV, Ivan ; NAIR, Indu ; FERRERI, Kevin ; AL-ABDULLAH, Ismail H ; MULLEN, Yoko ; KANDEEL, Fouad ; AVAKIAN-MANSOORIAN, Alina ; RAWSON, Jeffrey ; OMORI, Keiko ; ITO, Taihei ; VALIENTE, Luis ; IGLESIAS-MEZA, Itzia ; ORR, Chris ; SHIANG, Keh-Dong</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c434t-e4a780d984f2f18fff753151fd5741bd122c730fcfa1a4a53c0376b526fb4b1a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Apoptosis</topic><topic>Biological and medical sciences</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Fundamental immunology</topic><topic>Glucagon - metabolism</topic><topic>Humans</topic><topic>Insulin - metabolism</topic><topic>Insulin Secretion</topic><topic>Insulin-Secreting Cells - metabolism</topic><topic>Islets of Langerhans - cytology</topic><topic>Islets of Langerhans - physiology</topic><topic>Medical sciences</topic><topic>Pancreatic Polypeptide - metabolism</topic><topic>Somatostatin - metabolism</topic><topic>Surgery (general aspects). Transplantations, organ and tissue grafts. Graft diseases</topic><topic>Tissue, organ and graft immunology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>TODOROV, Ivan</creatorcontrib><creatorcontrib>NAIR, Indu</creatorcontrib><creatorcontrib>FERRERI, Kevin</creatorcontrib><creatorcontrib>AL-ABDULLAH, Ismail H</creatorcontrib><creatorcontrib>MULLEN, Yoko</creatorcontrib><creatorcontrib>KANDEEL, Fouad</creatorcontrib><creatorcontrib>AVAKIAN-MANSOORIAN, Alina</creatorcontrib><creatorcontrib>RAWSON, Jeffrey</creatorcontrib><creatorcontrib>OMORI, Keiko</creatorcontrib><creatorcontrib>ITO, Taihei</creatorcontrib><creatorcontrib>VALIENTE, Luis</creatorcontrib><creatorcontrib>IGLESIAS-MEZA, Itzia</creatorcontrib><creatorcontrib>ORR, Chris</creatorcontrib><creatorcontrib>SHIANG, Keh-Dong</creatorcontrib><collection>Pascal-Francis</collection><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><collection>PubMed Central (Full Participant titles)</collection><jtitle>Transplantation</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>TODOROV, Ivan</au><au>NAIR, Indu</au><au>FERRERI, Kevin</au><au>AL-ABDULLAH, Ismail H</au><au>MULLEN, Yoko</au><au>KANDEEL, Fouad</au><au>AVAKIAN-MANSOORIAN, Alina</au><au>RAWSON, Jeffrey</au><au>OMORI, Keiko</au><au>ITO, Taihei</au><au>VALIENTE, Luis</au><au>IGLESIAS-MEZA, Itzia</au><au>ORR, Chris</au><au>SHIANG, Keh-Dong</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Quantitative Assessment of β-Cell Apoptosis and Cell Composition of Isolated, Undisrupted Human Islets by Laser Scanning Cytometry</atitle><jtitle>Transplantation</jtitle><addtitle>Transplantation</addtitle><date>2010-10-27</date><risdate>2010</risdate><volume>90</volume><issue>8</issue><spage>836</spage><epage>842</epage><pages>836-842</pages><issn>0041-1337</issn><eissn>1534-6080</eissn><coden>TRPLAU</coden><abstract>Assays for assessing human islet cell quality, which provide results before transplantation, would be beneficial to improve the outcomes for islet transplantation therapy. Parameters such as percent β-cell apoptosis and cell composition are found to vary markedly between different islet preparations and may serve as markers of islet quality. We have developed fluorescence-based assays using laser scanning cytometry for assessing β-cell apoptosis and islet cell composition on serial sections of intact isolated islets.
Isolated human islets were fixed in formalin and embedded in paraffin. Serial sections were immunostained for the pancreatic hormones and acinar and ductal cell markers. DNA fragmentation was used to label apoptotic cells. Stained cells were quantified using an iCys laser scanning cytometer.
Islet preparations from 102 human pancreatic islet isolations were analyzed. For the whole set of islet preparations, we found a mean islet cell composition of 54.5%±1.2% insulin-positive, 33.9%±1.2% glucagon, 12.1%±0.7% somatostatin, and 1.5%±0.2% pancreatic polypeptide-positive cells. The apoptotic β cells were 2.85%±0.4% with a range of 0.27% to 18.3%. The percentage of apoptotic β cells correlated well (P<0.0001, n=59) with results obtained in vivo by transplantation of the corresponding islets in diabetic NODscid mice.
The analysis of whole, nondissociated islets for cell composition and β-cell apoptosis using laser scanning cytometry gives reliable and reproducible results and could be performed both before islet transplantation and on preserved cell blocks at any time in future. Thus, they can be a powerful tool for islet quality assessment.</abstract><cop>Hagerstown, MD</cop><pub>Lippincott Williams & Wilkins</pub><pmid>20697327</pmid><doi>10.1097/TP.0b013e3181f1db5d</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Transplantation, 2010-10, Vol.90 (8), p.836-842 |
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| source | MEDLINE; Journals@Ovid Complete |
| subjects | Apoptosis Biological and medical sciences Fundamental and applied biological sciences. Psychology Fundamental immunology Glucagon - metabolism Humans Insulin - metabolism Insulin Secretion Insulin-Secreting Cells - metabolism Islets of Langerhans - cytology Islets of Langerhans - physiology Medical sciences Pancreatic Polypeptide - metabolism Somatostatin - metabolism Surgery (general aspects). Transplantations, organ and tissue grafts. Graft diseases Tissue, organ and graft immunology |
| title | Quantitative Assessment of β-Cell Apoptosis and Cell Composition of Isolated, Undisrupted Human Islets by Laser Scanning Cytometry |
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