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OP-142 Gene Expression Profile of Nonfunctional Human Pancreatic Islets: Predictors of Transplant Failure?
Islet culture has become a standard part of most successful protocols for clinical islet transplantation. To date, however, islets are transplanted based on crude measures of viability, purity and in vitro insulin production without adequate prior assessment of the potential for in vivo function. Th...
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| Published in: | Transplantation proceedings 2005-10, Vol.37 (8), p.3441-3443 |
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| container_title | Transplantation proceedings |
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| creator | Sabek, O.M. Marshall, D.R. Minoru, O. Fraga, D.W. Gaber, A.O. |
| description | Islet culture has become a standard part of most successful protocols for clinical islet transplantation. To date, however, islets are transplanted based on crude measures of viability, purity and in vitro insulin production without adequate prior assessment of the potential for in vivo function. The purpose of this study was to define the gene expression profiles of human islets associated with in vivo function using a nonimmune NOD-scid mouse model. Human islets from eight isolations were maintained in culture for 7 to 14 days in Memphis serum-free media until transplanted. The RNA was extracted from 10,000 IEQ using RNASTAT-60. The gene expression profiles were analyzed using high-density Affymetrix U133A GeneChips and Genespring software. An aliquot of 2000 IEQ from each islet preparation was also transplanted into NOD-scid animals (n = 5) for in vivo function assessments. Islet function was assessed by measurements of human C-peptide at days 7 and 14 posttransplant. Human C-peptide levels were determined by radioimmunoassay. Gene analysis of nonfunction islets (4 of 8 islet preparations) showed high relative levels of expression of proinflammatory genes and low relative levels of genes directed toward insulin processing and secretion as well as islet integrity. Overexpression of hypoxia and proinflammatory genes may result in reduced insulin secretion and lead to islet destruction posttransplantation. Identifying and validating those genes could allow the development of a potency assay for human transplantation that would be very useful for screening human islet preparations before clinical transplant. |
| doi_str_mv | 10.1016/j.transproceed.2005.09.054 |
| format | article |
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To date, however, islets are transplanted based on crude measures of viability, purity and in vitro insulin production without adequate prior assessment of the potential for in vivo function. The purpose of this study was to define the gene expression profiles of human islets associated with in vivo function using a nonimmune NOD-scid mouse model. Human islets from eight isolations were maintained in culture for 7 to 14 days in Memphis serum-free media until transplanted. The RNA was extracted from 10,000 IEQ using RNASTAT-60. The gene expression profiles were analyzed using high-density Affymetrix U133A GeneChips and Genespring software. An aliquot of 2000 IEQ from each islet preparation was also transplanted into NOD-scid animals (n = 5) for in vivo function assessments. Islet function was assessed by measurements of human C-peptide at days 7 and 14 posttransplant. Human C-peptide levels were determined by radioimmunoassay. Gene analysis of nonfunction islets (4 of 8 islet preparations) showed high relative levels of expression of proinflammatory genes and low relative levels of genes directed toward insulin processing and secretion as well as islet integrity. Overexpression of hypoxia and proinflammatory genes may result in reduced insulin secretion and lead to islet destruction posttransplantation. Identifying and validating those genes could allow the development of a potency assay for human transplantation that would be very useful for screening human islet preparations before clinical transplant.</description><identifier>ISSN: 0041-1345</identifier><identifier>EISSN: 1873-2623</identifier><identifier>DOI: 10.1016/j.transproceed.2005.09.054</identifier><identifier>PMID: 16298622</identifier><identifier>CODEN: TRPPA8</identifier><language>eng</language><publisher>New York, NY: Elsevier Inc</publisher><subject>Adult ; Animals ; Biological and medical sciences ; C-Peptide - blood ; Child, Preschool ; Female ; Fundamental and applied biological sciences. Psychology ; Fundamental immunology ; Gene Expression Profiling ; Humans ; Insulin-Like Growth Factor Binding Protein 5 - genetics ; Islets of Langerhans - cytology ; Islets of Langerhans Transplantation - physiology ; Male ; Medical sciences ; Mice ; Mice, Inbred NOD ; Surgery (general aspects). 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To date, however, islets are transplanted based on crude measures of viability, purity and in vitro insulin production without adequate prior assessment of the potential for in vivo function. The purpose of this study was to define the gene expression profiles of human islets associated with in vivo function using a nonimmune NOD-scid mouse model. Human islets from eight isolations were maintained in culture for 7 to 14 days in Memphis serum-free media until transplanted. The RNA was extracted from 10,000 IEQ using RNASTAT-60. The gene expression profiles were analyzed using high-density Affymetrix U133A GeneChips and Genespring software. An aliquot of 2000 IEQ from each islet preparation was also transplanted into NOD-scid animals (n = 5) for in vivo function assessments. Islet function was assessed by measurements of human C-peptide at days 7 and 14 posttransplant. Human C-peptide levels were determined by radioimmunoassay. Gene analysis of nonfunction islets (4 of 8 islet preparations) showed high relative levels of expression of proinflammatory genes and low relative levels of genes directed toward insulin processing and secretion as well as islet integrity. Overexpression of hypoxia and proinflammatory genes may result in reduced insulin secretion and lead to islet destruction posttransplantation. Identifying and validating those genes could allow the development of a potency assay for human transplantation that would be very useful for screening human islet preparations before clinical transplant.</description><subject>Adult</subject><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>C-Peptide - blood</subject><subject>Child, Preschool</subject><subject>Female</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Fundamental immunology</subject><subject>Gene Expression Profiling</subject><subject>Humans</subject><subject>Insulin-Like Growth Factor Binding Protein 5 - genetics</subject><subject>Islets of Langerhans - cytology</subject><subject>Islets of Langerhans Transplantation - physiology</subject><subject>Male</subject><subject>Medical sciences</subject><subject>Mice</subject><subject>Mice, Inbred NOD</subject><subject>Surgery (general aspects). Transplantations, organ and tissue grafts. Graft diseases</subject><subject>Tissue, organ and graft immunology</subject><subject>Transplantation, Heterologous</subject><subject>Vascular Endothelial Growth Factor A - genetics</subject><issn>0041-1345</issn><issn>1873-2623</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2005</creationdate><recordtype>article</recordtype><recordid>eNqNkM2KFDEUhYMoTjv6ClIIuqsyv5XUbETmHwZnFuM6pG_dQJrqSpukBn37SduNunQVkvudm8NHyAdGO0ZZ_3nTleTmvEsREMeOU6o6OnRUyRdkxYwWLe-5eElWlErWMiHVCXmT84bWO5fiNTlhPR9Mz_mKbO4fWiZ5c40zNpc_dwlzDnFuHlL0YcIm-uZbnP0yQ6nPbmpulq2rYzdDQlcCNLd5wpLPagLHACWmvA89_m44ubk0Vy5MS8Ivb8kr76aM747nKfl-dfl4ftPe3V_fnn-9a0FSU9oRATWotZLOGafFWmuhlZTgDUhGNepe985xDoZ6UL4XnEoBg1v3aqSDF6fk02FvFfRjwVzsNmTAqZbBuGTbG8OZkbqCZwcQUsw5obe7FLYu_bKM2r1pu7H_mrZ705YOtpqu4ffHX5b1ts7-RI9qK_DxCLgMbvJ1EYT8l9OCKi733MWBw-rkKWCyGQLOUG0mhGLHGP6nzzOut6Pj</recordid><startdate>20051001</startdate><enddate>20051001</enddate><creator>Sabek, O.M.</creator><creator>Marshall, D.R.</creator><creator>Minoru, O.</creator><creator>Fraga, D.W.</creator><creator>Gaber, A.O.</creator><general>Elsevier Inc</general><general>Elsevier Science</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></search><sort><creationdate>20051001</creationdate><title>OP-142 Gene Expression Profile of Nonfunctional Human Pancreatic Islets: Predictors of Transplant Failure?</title><author>Sabek, O.M. ; Marshall, D.R. ; Minoru, O. ; Fraga, D.W. ; Gaber, A.O.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c408t-dece7c5b54aa8a73b7737544cf8c4107e7676aa22c80fc5f632043c9ab65d09f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2005</creationdate><topic>Adult</topic><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>C-Peptide - blood</topic><topic>Child, Preschool</topic><topic>Female</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Fundamental immunology</topic><topic>Gene Expression Profiling</topic><topic>Humans</topic><topic>Insulin-Like Growth Factor Binding Protein 5 - genetics</topic><topic>Islets of Langerhans - cytology</topic><topic>Islets of Langerhans Transplantation - physiology</topic><topic>Male</topic><topic>Medical sciences</topic><topic>Mice</topic><topic>Mice, Inbred NOD</topic><topic>Surgery (general aspects). Transplantations, organ and tissue grafts. Graft diseases</topic><topic>Tissue, organ and graft immunology</topic><topic>Transplantation, Heterologous</topic><topic>Vascular Endothelial Growth Factor A - genetics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Sabek, O.M.</creatorcontrib><creatorcontrib>Marshall, D.R.</creatorcontrib><creatorcontrib>Minoru, O.</creatorcontrib><creatorcontrib>Fraga, D.W.</creatorcontrib><creatorcontrib>Gaber, A.O.</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><jtitle>Transplantation proceedings</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Sabek, O.M.</au><au>Marshall, D.R.</au><au>Minoru, O.</au><au>Fraga, D.W.</au><au>Gaber, A.O.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>OP-142 Gene Expression Profile of Nonfunctional Human Pancreatic Islets: Predictors of Transplant Failure?</atitle><jtitle>Transplantation proceedings</jtitle><addtitle>Transplant Proc</addtitle><date>2005-10-01</date><risdate>2005</risdate><volume>37</volume><issue>8</issue><spage>3441</spage><epage>3443</epage><pages>3441-3443</pages><issn>0041-1345</issn><eissn>1873-2623</eissn><coden>TRPPA8</coden><notes>ObjectType-Article-1</notes><notes>SourceType-Scholarly Journals-1</notes><notes>ObjectType-Feature-2</notes><notes>content type line 23</notes><abstract>Islet culture has become a standard part of most successful protocols for clinical islet transplantation. To date, however, islets are transplanted based on crude measures of viability, purity and in vitro insulin production without adequate prior assessment of the potential for in vivo function. The purpose of this study was to define the gene expression profiles of human islets associated with in vivo function using a nonimmune NOD-scid mouse model. Human islets from eight isolations were maintained in culture for 7 to 14 days in Memphis serum-free media until transplanted. The RNA was extracted from 10,000 IEQ using RNASTAT-60. The gene expression profiles were analyzed using high-density Affymetrix U133A GeneChips and Genespring software. An aliquot of 2000 IEQ from each islet preparation was also transplanted into NOD-scid animals (n = 5) for in vivo function assessments. Islet function was assessed by measurements of human C-peptide at days 7 and 14 posttransplant. Human C-peptide levels were determined by radioimmunoassay. Gene analysis of nonfunction islets (4 of 8 islet preparations) showed high relative levels of expression of proinflammatory genes and low relative levels of genes directed toward insulin processing and secretion as well as islet integrity. Overexpression of hypoxia and proinflammatory genes may result in reduced insulin secretion and lead to islet destruction posttransplantation. Identifying and validating those genes could allow the development of a potency assay for human transplantation that would be very useful for screening human islet preparations before clinical transplant.</abstract><cop>New York, NY</cop><pub>Elsevier Inc</pub><pmid>16298622</pmid><doi>10.1016/j.transproceed.2005.09.054</doi><tpages>3</tpages></addata></record> |
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| subjects | Adult Animals Biological and medical sciences C-Peptide - blood Child, Preschool Female Fundamental and applied biological sciences. Psychology Fundamental immunology Gene Expression Profiling Humans Insulin-Like Growth Factor Binding Protein 5 - genetics Islets of Langerhans - cytology Islets of Langerhans Transplantation - physiology Male Medical sciences Mice Mice, Inbred NOD Surgery (general aspects). Transplantations, organ and tissue grafts. Graft diseases Tissue, organ and graft immunology Transplantation, Heterologous Vascular Endothelial Growth Factor A - genetics |
| title | OP-142 Gene Expression Profile of Nonfunctional Human Pancreatic Islets: Predictors of Transplant Failure? |
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