Adjustment of the Ratio Between Collagenase Class II and I Improves Islet Isolation Outcome
Previous studies have clarified the distinct roles of collagenase class I (ccI) and class II (ccII) in enzymatic release of islets from pancreatic tissue. The present study sought to enhance the limited knowledge about the optimal ratio between collagenase classes. Rat islets were isolated utilizing...
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| Veröffentlicht in: | Transplantation proceedings 2005-10, Vol.37 (8), p.3450-3451 |
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| creator | Brandhorst, D. Huettler, S. Alt, A. Raemsch-Guenther, N. Kurfuerst, M. Bretzel, R.G. Brandhorst, H. |
| description | Previous studies have clarified the distinct roles of collagenase class I (ccI) and class II (ccII) in enzymatic release of islets from pancreatic tissue. The present study sought to enhance the limited knowledge about the optimal ratio between collagenase classes.
Rat islets were isolated utilizing 0.4 DMC-U of neutral protease and 20 PZ-U of fractionated NB-1 collagenase recombined to obtain a ccII/I ratio of 0.5, 1.0, and 1.5. Quality control included assessment of yield (islet equivalents), trypan-blue exclusion, insulin release during static glucose incubation, and transplant function in diabetic nude mice. Data are expressed as mean values ± SEM.
Digestion time was only minimally influenced by different ccII/I ratios. The highest islet yield (
P < .05) was obtained using a ccII/I ratio of 1.0. Purity and glucose stimulation index were only marginally affected by different ccII/I ratios. A significant loss of islet viability after 24-hour culture (
P < .05) was observed only in islets isolated by means of a ccII/I ratio of 0.5 and 1.5 but not 1.0. Transplantation into diabetic nude mice revealed sustained islet graft function in all experimental groups.
The present study indicates that the ratio between ccII and ccI is of significant relevance for optimizing islet yield and viability. |
| doi_str_mv | 10.1016/j.transproceed.2005.09.079 |
| format | Article |
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Rat islets were isolated utilizing 0.4 DMC-U of neutral protease and 20 PZ-U of fractionated NB-1 collagenase recombined to obtain a ccII/I ratio of 0.5, 1.0, and 1.5. Quality control included assessment of yield (islet equivalents), trypan-blue exclusion, insulin release during static glucose incubation, and transplant function in diabetic nude mice. Data are expressed as mean values ± SEM.
Digestion time was only minimally influenced by different ccII/I ratios. The highest islet yield (
P < .05) was obtained using a ccII/I ratio of 1.0. Purity and glucose stimulation index were only marginally affected by different ccII/I ratios. A significant loss of islet viability after 24-hour culture (
P < .05) was observed only in islets isolated by means of a ccII/I ratio of 0.5 and 1.5 but not 1.0. Transplantation into diabetic nude mice revealed sustained islet graft function in all experimental groups.
The present study indicates that the ratio between ccII and ccI is of significant relevance for optimizing islet yield and viability.</description><identifier>ISSN: 0041-1345</identifier><identifier>EISSN: 1873-2623</identifier><identifier>DOI: 10.1016/j.transproceed.2005.09.079</identifier><identifier>PMID: 16298625</identifier><identifier>CODEN: TRPPA8</identifier><language>eng</language><publisher>New York, NY: Elsevier Inc</publisher><subject>Animals ; Biological and medical sciences ; Biomarkers - analysis ; Cell Separation - methods ; Collagenases - analysis ; Fundamental and applied biological sciences. Psychology ; Fundamental immunology ; Islets of Langerhans - cytology ; Islets of Langerhans - enzymology ; Medical sciences ; Rats ; Surgery (general aspects). Transplantations, organ and tissue grafts. Graft diseases ; Tissue, organ and graft immunology</subject><ispartof>Transplantation proceedings, 2005-10, Vol.37 (8), p.3450-3451</ispartof><rights>2005 Elsevier Inc.</rights><rights>2006 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c408t-9ffc996e42cef5a89765a74957d7050a1e5ed92c81cbc8fb72cd3fdbb6fb619e3</citedby><cites>FETCH-LOGICAL-c408t-9ffc996e42cef5a89765a74957d7050a1e5ed92c81cbc8fb72cd3fdbb6fb619e3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.transproceed.2005.09.079$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>309,310,314,780,784,789,790,3550,23930,23931,25140,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=17305245$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16298625$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Brandhorst, D.</creatorcontrib><creatorcontrib>Huettler, S.</creatorcontrib><creatorcontrib>Alt, A.</creatorcontrib><creatorcontrib>Raemsch-Guenther, N.</creatorcontrib><creatorcontrib>Kurfuerst, M.</creatorcontrib><creatorcontrib>Bretzel, R.G.</creatorcontrib><creatorcontrib>Brandhorst, H.</creatorcontrib><title>Adjustment of the Ratio Between Collagenase Class II and I Improves Islet Isolation Outcome</title><title>Transplantation proceedings</title><addtitle>Transplant Proc</addtitle><description>Previous studies have clarified the distinct roles of collagenase class I (ccI) and class II (ccII) in enzymatic release of islets from pancreatic tissue. The present study sought to enhance the limited knowledge about the optimal ratio between collagenase classes.
Rat islets were isolated utilizing 0.4 DMC-U of neutral protease and 20 PZ-U of fractionated NB-1 collagenase recombined to obtain a ccII/I ratio of 0.5, 1.0, and 1.5. Quality control included assessment of yield (islet equivalents), trypan-blue exclusion, insulin release during static glucose incubation, and transplant function in diabetic nude mice. Data are expressed as mean values ± SEM.
Digestion time was only minimally influenced by different ccII/I ratios. The highest islet yield (
P < .05) was obtained using a ccII/I ratio of 1.0. Purity and glucose stimulation index were only marginally affected by different ccII/I ratios. A significant loss of islet viability after 24-hour culture (
P < .05) was observed only in islets isolated by means of a ccII/I ratio of 0.5 and 1.5 but not 1.0. Transplantation into diabetic nude mice revealed sustained islet graft function in all experimental groups.
The present study indicates that the ratio between ccII and ccI is of significant relevance for optimizing islet yield and viability.</description><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Biomarkers - analysis</subject><subject>Cell Separation - methods</subject><subject>Collagenases - analysis</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Fundamental immunology</subject><subject>Islets of Langerhans - cytology</subject><subject>Islets of Langerhans - enzymology</subject><subject>Medical sciences</subject><subject>Rats</subject><subject>Surgery (general aspects). Transplantations, organ and tissue grafts. Graft diseases</subject><subject>Tissue, organ and graft immunology</subject><issn>0041-1345</issn><issn>1873-2623</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2005</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkE2P1SAUhonRONfRv2CIie5agRYK7mauX00mmcToygWhcNDetGXsoWP893Jzb9SlGwjwvIc3DyEvOKs54-r1oc6rW_BuTR4g1IIxWTNTs848IDuuu6YSSjQPyY6xlle8aeUFeYJ4YOUs2uYxueBKGK2E3JGvV-GwYZ5hyTRFmr8D_eTymOg15J8AC92naXLfYHEIdD85RNr31C2B9rSfS4V7KDc4QS5rmo7Rhd5u2acZnpJH0U0Iz877Jfny_t3n_cfq5vZDv7-6qXzLdK5MjN4YBa3wEKXTplPSda2RXeiYZI6DhGCE19wPXsehEz40MQyDioPiBppL8uo0t9T5sQFmO4_oofReIG1oldZca60K-OYE-jUhrhDt3TrObv1lObNHtfZg_1Vrj2otM7aoLeHn51-2YS5vf6JnlwV4eQYcejfFMsiP-JfrGiZFe-TenjgoTu5HWC36ERYPYVzBZxvS-D99fgMf7J9n</recordid><startdate>20051001</startdate><enddate>20051001</enddate><creator>Brandhorst, D.</creator><creator>Huettler, S.</creator><creator>Alt, A.</creator><creator>Raemsch-Guenther, N.</creator><creator>Kurfuerst, M.</creator><creator>Bretzel, R.G.</creator><creator>Brandhorst, H.</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>Adjustment of the Ratio Between Collagenase Class II and I Improves Islet Isolation Outcome</title><author>Brandhorst, D. ; Huettler, S. ; Alt, A. ; Raemsch-Guenther, N. ; Kurfuerst, M. ; Bretzel, R.G. ; Brandhorst, H.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c408t-9ffc996e42cef5a89765a74957d7050a1e5ed92c81cbc8fb72cd3fdbb6fb619e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2005</creationdate><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Biomarkers - analysis</topic><topic>Cell Separation - methods</topic><topic>Collagenases - analysis</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Fundamental immunology</topic><topic>Islets of Langerhans - cytology</topic><topic>Islets of Langerhans - enzymology</topic><topic>Medical sciences</topic><topic>Rats</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>Brandhorst, D.</creatorcontrib><creatorcontrib>Huettler, S.</creatorcontrib><creatorcontrib>Alt, A.</creatorcontrib><creatorcontrib>Raemsch-Guenther, N.</creatorcontrib><creatorcontrib>Kurfuerst, M.</creatorcontrib><creatorcontrib>Bretzel, R.G.</creatorcontrib><creatorcontrib>Brandhorst, H.</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>Brandhorst, D.</au><au>Huettler, S.</au><au>Alt, A.</au><au>Raemsch-Guenther, N.</au><au>Kurfuerst, M.</au><au>Bretzel, R.G.</au><au>Brandhorst, H.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Adjustment of the Ratio Between Collagenase Class II and I Improves Islet Isolation Outcome</atitle><jtitle>Transplantation proceedings</jtitle><addtitle>Transplant Proc</addtitle><date>2005-10-01</date><risdate>2005</risdate><volume>37</volume><issue>8</issue><spage>3450</spage><epage>3451</epage><pages>3450-3451</pages><issn>0041-1345</issn><eissn>1873-2623</eissn><coden>TRPPA8</coden><abstract>Previous studies have clarified the distinct roles of collagenase class I (ccI) and class II (ccII) in enzymatic release of islets from pancreatic tissue. The present study sought to enhance the limited knowledge about the optimal ratio between collagenase classes.
Rat islets were isolated utilizing 0.4 DMC-U of neutral protease and 20 PZ-U of fractionated NB-1 collagenase recombined to obtain a ccII/I ratio of 0.5, 1.0, and 1.5. Quality control included assessment of yield (islet equivalents), trypan-blue exclusion, insulin release during static glucose incubation, and transplant function in diabetic nude mice. Data are expressed as mean values ± SEM.
Digestion time was only minimally influenced by different ccII/I ratios. The highest islet yield (
P < .05) was obtained using a ccII/I ratio of 1.0. Purity and glucose stimulation index were only marginally affected by different ccII/I ratios. A significant loss of islet viability after 24-hour culture (
P < .05) was observed only in islets isolated by means of a ccII/I ratio of 0.5 and 1.5 but not 1.0. Transplantation into diabetic nude mice revealed sustained islet graft function in all experimental groups.
The present study indicates that the ratio between ccII and ccI is of significant relevance for optimizing islet yield and viability.</abstract><cop>New York, NY</cop><pub>Elsevier Inc</pub><pmid>16298625</pmid><doi>10.1016/j.transproceed.2005.09.079</doi><tpages>2</tpages></addata></record> |
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| subjects | Animals Biological and medical sciences Biomarkers - analysis Cell Separation - methods Collagenases - analysis Fundamental and applied biological sciences. Psychology Fundamental immunology Islets of Langerhans - cytology Islets of Langerhans - enzymology Medical sciences Rats Surgery (general aspects). Transplantations, organ and tissue grafts. Graft diseases Tissue, organ and graft immunology |
| title | Adjustment of the Ratio Between Collagenase Class II and I Improves Islet Isolation Outcome |
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