Monolayer-Forming Islet Cell Culture from Neonatal Pig Pancreas: Using Sequential Treatment with EDTA-Dispase and Monoiodoacetic Acid for Preparation and Purification
OHGAWARA, H., IWANAGA, T., YUI, R., NISHIJIMA, S. and HIRATA, Y. Monolayer-Forming Islet Cell Culture from Neonatal Pig Pancreas: Using Sequential Treatment with EDTA-Dispase and Monoiodoacetic Acid for Preparation and Purification. Tohoku J. exp. Med., 1987, 153 (4), 375-382 - A new method for the...
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| Veröffentlicht in: | The Tohoku Journal of Experimental Medicine 1987, Vol.153(4), pp.375-382 |
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| container_title | The Tohoku Journal of Experimental Medicine |
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| creator | OHGAWARA, HISAKO IWANAGA, TOSHIHIKO YUI, RYOGO NISHIJIMA, SACHIYO HIRATA, YUKIMASA |
| description | OHGAWARA, H., IWANAGA, T., YUI, R., NISHIJIMA, S. and HIRATA, Y. Monolayer-Forming Islet Cell Culture from Neonatal Pig Pancreas: Using Sequential Treatment with EDTA-Dispase and Monoiodoacetic Acid for Preparation and Purification. Tohoku J. exp. Med., 1987, 153 (4), 375-382 - A new method for the preparation and purification of a monolayer-forming islet cell culture from the neonatal pig pancreas is described. Single cell preparations of the pig pancreas were obtained by gently stirring the chopped pancreases in a medium containing sequential EDTA-dispase. Exposure of the cells to monoiodoacetic acid (10μm) and BSA-gradient resulted in improved preparation, highly enriched in the endocrine cells. The cells were maintained in tissue culture medium 199 containing 5.5mM D-glucose, with or without 0.1mM 3-isobutyl-1-methylxanthine, or 16.7mM D-glucose and 20% fetal calf serum. During a 6-day culture period, many small cell aggregates appeared in the media. The cell clusters attached to the bottom of the dish and formed monolayers. Provocative stimulation and radioimmuno assay for insulin showed the existence of numerous and viable B cells in the cell clusters. The other three types of islet cells were also demonstrated immunohistochemically in the cell cluster. It is concluded that this improved culsture technique provides a useful tool for morphological and biochemical studies of islet cells and a potential source of material for transplantation of B-cells from the pancreas. |
| doi_str_mv | 10.1620/tjem.153.375 |
| format | Article |
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Monolayer-Forming Islet Cell Culture from Neonatal Pig Pancreas: Using Sequential Treatment with EDTA-Dispase and Monoiodoacetic Acid for Preparation and Purification. Tohoku J. exp. Med., 1987, 153 (4), 375-382 - A new method for the preparation and purification of a monolayer-forming islet cell culture from the neonatal pig pancreas is described. Single cell preparations of the pig pancreas were obtained by gently stirring the chopped pancreases in a medium containing sequential EDTA-dispase. Exposure of the cells to monoiodoacetic acid (10μm) and BSA-gradient resulted in improved preparation, highly enriched in the endocrine cells. The cells were maintained in tissue culture medium 199 containing 5.5mM D-glucose, with or without 0.1mM 3-isobutyl-1-methylxanthine, or 16.7mM D-glucose and 20% fetal calf serum. During a 6-day culture period, many small cell aggregates appeared in the media. The cell clusters attached to the bottom of the dish and formed monolayers. Provocative stimulation and radioimmuno assay for insulin showed the existence of numerous and viable B cells in the cell clusters. The other three types of islet cells were also demonstrated immunohistochemically in the cell cluster. It is concluded that this improved culsture technique provides a useful tool for morphological and biochemical studies of islet cells and a potential source of material for transplantation of B-cells from the pancreas.</description><identifier>ISSN: 0040-8727</identifier><identifier>EISSN: 1349-3329</identifier><identifier>DOI: 10.1620/tjem.153.375</identifier><identifier>PMID: 2450412</identifier><language>eng</language><publisher>Japan: Tohoku University Medical Press</publisher><subject>1-Methyl-3-isobutylxanthine - pharmacology ; Animals ; Animals, Newborn ; Cell Aggregation ; Cell Separation - methods ; Cells, Cultured ; Edetic Acid ; EDTA-dispase ; Endopeptidases ; Fluorescent Antibody Technique ; Insulin - metabolism ; Insulin Secretion ; Iodoacetates ; Iodoacetic Acid ; Islets of Langerhans - cytology ; Islets of Langerhans - drug effects ; Islets of Langerhans - metabolism ; monoiodoacetic acid ; monolayer culture ; pig pancreas ; Swine</subject><ispartof>The Tohoku Journal of Experimental Medicine, 1987, Vol.153(4), pp.375-382</ispartof><rights>Tohoku University Medical Press</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c453t-d57f56db95b39187eb59b5514320aec19fa18f0a82876bb76460c456a0357153</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,1883,4024,27923,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/2450412$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>OHGAWARA, HISAKO</creatorcontrib><creatorcontrib>IWANAGA, TOSHIHIKO</creatorcontrib><creatorcontrib>YUI, RYOGO</creatorcontrib><creatorcontrib>NISHIJIMA, SACHIYO</creatorcontrib><creatorcontrib>HIRATA, YUKIMASA</creatorcontrib><title>Monolayer-Forming Islet Cell Culture from Neonatal Pig Pancreas: Using Sequential Treatment with EDTA-Dispase and Monoiodoacetic Acid for Preparation and Purification</title><title>The Tohoku Journal of Experimental Medicine</title><addtitle>Tohoku J. Exp. Med.</addtitle><description>OHGAWARA, H., IWANAGA, T., YUI, R., NISHIJIMA, S. and HIRATA, Y. Monolayer-Forming Islet Cell Culture from Neonatal Pig Pancreas: Using Sequential Treatment with EDTA-Dispase and Monoiodoacetic Acid for Preparation and Purification. Tohoku J. exp. Med., 1987, 153 (4), 375-382 - A new method for the preparation and purification of a monolayer-forming islet cell culture from the neonatal pig pancreas is described. Single cell preparations of the pig pancreas were obtained by gently stirring the chopped pancreases in a medium containing sequential EDTA-dispase. Exposure of the cells to monoiodoacetic acid (10μm) and BSA-gradient resulted in improved preparation, highly enriched in the endocrine cells. The cells were maintained in tissue culture medium 199 containing 5.5mM D-glucose, with or without 0.1mM 3-isobutyl-1-methylxanthine, or 16.7mM D-glucose and 20% fetal calf serum. During a 6-day culture period, many small cell aggregates appeared in the media. The cell clusters attached to the bottom of the dish and formed monolayers. Provocative stimulation and radioimmuno assay for insulin showed the existence of numerous and viable B cells in the cell clusters. The other three types of islet cells were also demonstrated immunohistochemically in the cell cluster. It is concluded that this improved culsture technique provides a useful tool for morphological and biochemical studies of islet cells and a potential source of material for transplantation of B-cells from the pancreas.</description><subject>1-Methyl-3-isobutylxanthine - pharmacology</subject><subject>Animals</subject><subject>Animals, Newborn</subject><subject>Cell Aggregation</subject><subject>Cell Separation - methods</subject><subject>Cells, Cultured</subject><subject>Edetic Acid</subject><subject>EDTA-dispase</subject><subject>Endopeptidases</subject><subject>Fluorescent Antibody Technique</subject><subject>Insulin - metabolism</subject><subject>Insulin Secretion</subject><subject>Iodoacetates</subject><subject>Iodoacetic Acid</subject><subject>Islets of Langerhans - cytology</subject><subject>Islets of Langerhans - drug effects</subject><subject>Islets of Langerhans - metabolism</subject><subject>monoiodoacetic acid</subject><subject>monolayer culture</subject><subject>pig pancreas</subject><subject>Swine</subject><issn>0040-8727</issn><issn>1349-3329</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1987</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNo9kUGP0zAQhS0EWsrCjSuST5xIsWM7TrhV3S2stEAlyjmaOJOuqyQutiO0f4jfidNWvdjyvDffyPMIec_Zkhc5-xwPOCy5Ekuh1Quy4EJWmRB59ZIsGJMsK3WuX5M3IRwYE5Lp4obc5FIxyfMF-ffdja6HZ_TZxvnBjnv6EHqMdI19T9dTHyePtPNuoD_QjRChp1u7p1sYjUcIX-jvMDf9wj8TjtEmeZfqcUgP-tfGJ3p_t1tldzYcISCFsaXzROtaBwajNXRlbEs75-nW4xE8ROvGk287edtZcyq8Ja866AO-u9y3ZLe5362_ZY8_vz6sV4-ZkUrErFW6U0XbVKoRFS81NqpqlOJS5AzQ8KoDXnYMyrzURdPoQhYsdRbAhNJphbfk4xl79C79J8R6sMGkTcCIbgq11lUpqyJPxk9no_EuBI9dffR2AP9cc1bPqdRzKnVC1imVZP9w4U7NgO3VfIkh6ZuzfggR9njVwacN9XiC8SpRZ6C8nAl8NZgn8DWO4j8QtKMh</recordid><startdate>1987</startdate><enddate>1987</enddate><creator>OHGAWARA, HISAKO</creator><creator>IWANAGA, TOSHIHIKO</creator><creator>YUI, RYOGO</creator><creator>NISHIJIMA, SACHIYO</creator><creator>HIRATA, YUKIMASA</creator><general>Tohoku University Medical Press</general><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>1987</creationdate><title>Monolayer-Forming Islet Cell Culture from Neonatal Pig Pancreas: Using Sequential Treatment with EDTA-Dispase and Monoiodoacetic Acid for Preparation and Purification</title><author>OHGAWARA, HISAKO ; IWANAGA, TOSHIHIKO ; YUI, RYOGO ; NISHIJIMA, SACHIYO ; HIRATA, YUKIMASA</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c453t-d57f56db95b39187eb59b5514320aec19fa18f0a82876bb76460c456a0357153</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1987</creationdate><topic>1-Methyl-3-isobutylxanthine - pharmacology</topic><topic>Animals</topic><topic>Animals, Newborn</topic><topic>Cell Aggregation</topic><topic>Cell Separation - methods</topic><topic>Cells, Cultured</topic><topic>Edetic Acid</topic><topic>EDTA-dispase</topic><topic>Endopeptidases</topic><topic>Fluorescent Antibody Technique</topic><topic>Insulin - metabolism</topic><topic>Insulin Secretion</topic><topic>Iodoacetates</topic><topic>Iodoacetic Acid</topic><topic>Islets of Langerhans - cytology</topic><topic>Islets of Langerhans - drug effects</topic><topic>Islets of Langerhans - metabolism</topic><topic>monoiodoacetic acid</topic><topic>monolayer culture</topic><topic>pig pancreas</topic><topic>Swine</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>OHGAWARA, HISAKO</creatorcontrib><creatorcontrib>IWANAGA, TOSHIHIKO</creatorcontrib><creatorcontrib>YUI, RYOGO</creatorcontrib><creatorcontrib>NISHIJIMA, SACHIYO</creatorcontrib><creatorcontrib>HIRATA, YUKIMASA</creatorcontrib><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>The Tohoku Journal of Experimental Medicine</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>OHGAWARA, HISAKO</au><au>IWANAGA, TOSHIHIKO</au><au>YUI, RYOGO</au><au>NISHIJIMA, SACHIYO</au><au>HIRATA, YUKIMASA</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Monolayer-Forming Islet Cell Culture from Neonatal Pig Pancreas: Using Sequential Treatment with EDTA-Dispase and Monoiodoacetic Acid for Preparation and Purification</atitle><jtitle>The Tohoku Journal of Experimental Medicine</jtitle><addtitle>Tohoku J. Exp. Med.</addtitle><date>1987</date><risdate>1987</risdate><volume>153</volume><issue>4</issue><spage>375</spage><epage>382</epage><pages>375-382</pages><issn>0040-8727</issn><eissn>1349-3329</eissn><abstract>OHGAWARA, H., IWANAGA, T., YUI, R., NISHIJIMA, S. and HIRATA, Y. Monolayer-Forming Islet Cell Culture from Neonatal Pig Pancreas: Using Sequential Treatment with EDTA-Dispase and Monoiodoacetic Acid for Preparation and Purification. Tohoku J. exp. Med., 1987, 153 (4), 375-382 - A new method for the preparation and purification of a monolayer-forming islet cell culture from the neonatal pig pancreas is described. Single cell preparations of the pig pancreas were obtained by gently stirring the chopped pancreases in a medium containing sequential EDTA-dispase. Exposure of the cells to monoiodoacetic acid (10μm) and BSA-gradient resulted in improved preparation, highly enriched in the endocrine cells. The cells were maintained in tissue culture medium 199 containing 5.5mM D-glucose, with or without 0.1mM 3-isobutyl-1-methylxanthine, or 16.7mM D-glucose and 20% fetal calf serum. During a 6-day culture period, many small cell aggregates appeared in the media. The cell clusters attached to the bottom of the dish and formed monolayers. Provocative stimulation and radioimmuno assay for insulin showed the existence of numerous and viable B cells in the cell clusters. The other three types of islet cells were also demonstrated immunohistochemically in the cell cluster. It is concluded that this improved culsture technique provides a useful tool for morphological and biochemical studies of islet cells and a potential source of material for transplantation of B-cells from the pancreas.</abstract><cop>Japan</cop><pub>Tohoku University Medical Press</pub><pmid>2450412</pmid><doi>10.1620/tjem.153.375</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record> |
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| source | J-STAGE Free; MEDLINE; Freely Accessible Japanese Titles; EZB-FREE-00999 freely available EZB journals |
| subjects | 1-Methyl-3-isobutylxanthine - pharmacology Animals Animals, Newborn Cell Aggregation Cell Separation - methods Cells, Cultured Edetic Acid EDTA-dispase Endopeptidases Fluorescent Antibody Technique Insulin - metabolism Insulin Secretion Iodoacetates Iodoacetic Acid Islets of Langerhans - cytology Islets of Langerhans - drug effects Islets of Langerhans - metabolism monoiodoacetic acid monolayer culture pig pancreas Swine |
| title | Monolayer-Forming Islet Cell Culture from Neonatal Pig Pancreas: Using Sequential Treatment with EDTA-Dispase and Monoiodoacetic Acid for Preparation and Purification |
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