Analysis of blood coagulation in mice: pre-analytical conditions and evaluation of a home-made assay for thrombin-antithrombin complexes
<p>Abstract</p> <p>Background</p> <p>The use of mouse models for the study of thrombotic disorders has gained increasing importance. Methods for measurement of coagulation activation in mice are, however, scarce. The primary aim of this study was to develop a specific m...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2005-08-01
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| Series: | Thrombosis Journal |
| Online Access: | http://www.thrombosisjournal.com/content/3/1/12 |
| _version_ | 1818021796069244928 |
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| author | Meijers Joost CM Spronk Henri MH Timmerman Janneke J Reitsma Pieter H van Oerle René Sommeijer Dirkje W ten Cate Hugo |
| author_facet | Meijers Joost CM Spronk Henri MH Timmerman Janneke J Reitsma Pieter H van Oerle René Sommeijer Dirkje W ten Cate Hugo |
| author_sort | Meijers Joost CM |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>The use of mouse models for the study of thrombotic disorders has gained increasing importance. Methods for measurement of coagulation activation in mice are, however, scarce. The primary aim of this study was to develop a specific mouse thrombin-antithrombin (TAT) ELISA for measurement of coagulation activation and to compare it with two commercially available assays for human TAT complexes. In addition, we aimed to improve methods for mouse plasma anticoagulation and preparation.</p> <p>Methods and results</p> <p>First, for the measurement of TAT-complexes in plasma a mouse specific TAT-ELISA was developed using rabbit polyclonal antibodies raised against mouse thrombin and rat antithrombin, respectively. This ELISA detected an increase in TAT levels in a mouse model of endotoxemia. Two commercial human TAT ELISAs appeared to be less specific for mouse thrombin-rat antithrombin complexes.</p> <p>Second, to prevent clotting of mouse blood sodium citrate was either mixed with blood during collection in a syringe or was injected intravenously immediately prior to blood collection. Intravenous sodium citrate completely inhibited blood coagulation resulting in plasma with consistently low TAT levels. Sodium citrate mixed with blood during collection resulted in increased TAT levels in 4 out of 16 plasma samples. Third, heparinase was added to plasma samples after in vivo injection of different heparin doses to test its neutralizing effect. Heparinase neutralized up to a 20 U of heparin/mouse and resulted in accurate APTT and factor VIII determinations.</p> <p>Conclusion</p> <p>These procedures and reagents for plasma preparation and coagulation testing will improve studies on thrombotic disorders in mice.</p> |
| first_indexed | 2024-04-14T08:22:45Z |
| format | Article |
| id | doaj.art-0ab9c447da4946968d4edb35812e3815 |
| institution | Directory Open Access Journal |
| issn | 1477-9560 |
| language | English |
| last_indexed | 2024-04-14T08:22:45Z |
| publishDate | 2005-08-01 |
| publisher | BMC |
| record_format | Article |
| series | Thrombosis Journal |
| spelling | doaj.art-0ab9c447da4946968d4edb35812e38152022-12-22T02:04:09ZengBMCThrombosis Journal1477-95602005-08-01311210.1186/1477-9560-3-12Analysis of blood coagulation in mice: pre-analytical conditions and evaluation of a home-made assay for thrombin-antithrombin complexesMeijers Joost CMSpronk Henri MHTimmerman Janneke JReitsma Pieter Hvan Oerle RenéSommeijer Dirkje Wten Cate Hugo<p>Abstract</p> <p>Background</p> <p>The use of mouse models for the study of thrombotic disorders has gained increasing importance. Methods for measurement of coagulation activation in mice are, however, scarce. The primary aim of this study was to develop a specific mouse thrombin-antithrombin (TAT) ELISA for measurement of coagulation activation and to compare it with two commercially available assays for human TAT complexes. In addition, we aimed to improve methods for mouse plasma anticoagulation and preparation.</p> <p>Methods and results</p> <p>First, for the measurement of TAT-complexes in plasma a mouse specific TAT-ELISA was developed using rabbit polyclonal antibodies raised against mouse thrombin and rat antithrombin, respectively. This ELISA detected an increase in TAT levels in a mouse model of endotoxemia. Two commercial human TAT ELISAs appeared to be less specific for mouse thrombin-rat antithrombin complexes.</p> <p>Second, to prevent clotting of mouse blood sodium citrate was either mixed with blood during collection in a syringe or was injected intravenously immediately prior to blood collection. Intravenous sodium citrate completely inhibited blood coagulation resulting in plasma with consistently low TAT levels. Sodium citrate mixed with blood during collection resulted in increased TAT levels in 4 out of 16 plasma samples. Third, heparinase was added to plasma samples after in vivo injection of different heparin doses to test its neutralizing effect. Heparinase neutralized up to a 20 U of heparin/mouse and resulted in accurate APTT and factor VIII determinations.</p> <p>Conclusion</p> <p>These procedures and reagents for plasma preparation and coagulation testing will improve studies on thrombotic disorders in mice.</p>http://www.thrombosisjournal.com/content/3/1/12 |
| spellingShingle | Meijers Joost CM Spronk Henri MH Timmerman Janneke J Reitsma Pieter H van Oerle René Sommeijer Dirkje W ten Cate Hugo Analysis of blood coagulation in mice: pre-analytical conditions and evaluation of a home-made assay for thrombin-antithrombin complexes Thrombosis Journal |
| title | Analysis of blood coagulation in mice: pre-analytical conditions and evaluation of a home-made assay for thrombin-antithrombin complexes |
| title_full | Analysis of blood coagulation in mice: pre-analytical conditions and evaluation of a home-made assay for thrombin-antithrombin complexes |
| title_fullStr | Analysis of blood coagulation in mice: pre-analytical conditions and evaluation of a home-made assay for thrombin-antithrombin complexes |
| title_full_unstemmed | Analysis of blood coagulation in mice: pre-analytical conditions and evaluation of a home-made assay for thrombin-antithrombin complexes |
| title_short | Analysis of blood coagulation in mice: pre-analytical conditions and evaluation of a home-made assay for thrombin-antithrombin complexes |
| title_sort | analysis of blood coagulation in mice pre analytical conditions and evaluation of a home made assay for thrombin antithrombin complexes |
| url | http://www.thrombosisjournal.com/content/3/1/12 |
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