Journal of Thrombosis and Haemostasis, 12: 1940–1942
DOI: 10.1111/jth.12700
RECOMMENDATIONS AND GUIDELINES
Influence of genetic background on bleeding phenotype in the tail-tip bleeding model and recommendations for standardization: communication from the SSC of the ISTH € LER,* M. SCHUSTER,* E.-M. MUCHITSCH* and A . S C H I V I Z , * D . M A G I R R , † P . L E I D E N M UH ¨ W. HOLLRIEGL,* FOR THE SUBCOMMITTEE ON ANIMAL MODELS *Baxter Innovations GmbH; and †Medical Statistics Section, Center for Medical Statistics, Informatics and Intelligent Systems, Medical University of Vienna, Vienna, Austria
€ llriegl W, for the Subcommittee on Animal Models. To cite this article: Schiviz A, Magirr D, Leidenm€ uhler P, Schuster M, Muchitsch EM, Ho Influence of genetic background on bleeding phenotype in the tail-tip bleeding model and recommendations for standardization: communication from the SSC of the ISTH. J Thromb Haemost 2014; 12:1940–2.
The murine tail-tip bleeding model is widely used to measure bleeding time, blood loss, and survival after treatment [1–4]. Because of the considerable variation in data from these studies and thus the need for large sample sizes per group, and the lack of comparability of results from different laboratories, research groups have tried to standardize the model. In 2010, the SSC of the ISTH proposed standardized methods for determining tail bleeding time in mice. The SSC report standardized the tail clipping by endorsing the use of a warming chamber to evenly warm the anesthetized mouse, and assessing the influence of anesthesia and systolic blood pressure on blood loss [5]. We sought to test the ability of the standardized method to detect differences in bleeding phenotypes of wild-type mouse strains, which may show different bleeding phenotypes [6]. The relevance of the genetic background of inbred mouse strains or transgenic mice on phenotype has been described in various research fields. Mice may differ in physiologic parameters [7] or in response to triggers. Polymorphisms exist for substrains of the same inbred strains [8]. Genetic background also influences pathways that regulate thrombosis and hemostasis: differences were observed in global bleeding and thrombosis assays [9], coagulation and fibrinolytic factors [10], and susceptibility to treatment [11].
Correspondence: Eva-Maria Muchitsch, Baxter Innovations GmbH, Vienna DC Tower, Donau-City-Straße 7, 1220 Vienna, Austria. Tel.: +43 1 20100 247 2626; fax: +43 1 20100 557. E-mail: [email protected] Received 1 October 2013 Manuscript handled by: S. Eichinger Final decision: P. H. Reitsma, 28 July 2014
To investigate the ability of the tail-tip bleeding assay to detect the influence of genetic background on bleeding phenotype, we used hemophilic mice on different backgrounds, i.e. coagulation factor VIII (exon 17) knockout (FVIII / ) [12] mice on a BALB/c, C57BL/6, or mixed background, along with different substrains of wild-type C57BL/6 or BALB/c mice (Table 1). The mice were anesthetized with 100 mg kg-1 ketamine and 10 mg kg-1 xylazine and placed in a warming chamber. Two millimeters of the tail tip were clipped with a guillotine [4], the tails were immersed in warm saline (2 mL, 35 2 °C), and blood was collected over a period of 60 min; anesthesia was maintained as required by subcutaneous supplementation with one-third of the starting dose. Blood loss was determined gravimetrically. At the end of this observation period, the mice were removed from the chamber and humanely killed by cervical dislocation. Pairwise tests of equality of variance were performed with a bootstrap approach; the level of statistical significance was set to P = 0.05. C.FVIII / mice showed markedly lower median blood loss (365.7 mg) than B6.FVIII / (965.2 mg) and B6; FVIII / (890.4 mg) mice. For all three strains, median blood loss was lower in females than in males, with the largest sex difference (634 mg vs. 954 mg for females vs. males) and the highest interindividual variation being observed in B6;FVIII / mice (Fig. 1). In 129S1/SvmJ mice and the four substrains of BALB/c mice, median blood loss (23.5 mg and 10.0–19.0 mg, respectively) and interanimal variability were low. In contrast, median blood loss in C57BL/6 substrains varied from 36.0 mg (C57BL/6BomTac) to 469.5 mg (C57BL/6JCrl). Both the intersubstrain and interindividual ranges of blood loss (Fig. 1) and sex differences in median blood loss were high in C57BL/6 mice (e.g. 228 mg vs. 870 mg in C57BL/ 6JCrl and 76 mg vs. 765 mg in C57BL/6OlaHsd for females vs. males). © 2014 International Society on Thrombosis and Haemostasis
Mouse strains in the tail-tip bleeding model 1941 Table 1 Mouse strains Strain FVIII
Substrain C.FVIII (C.129S4-F8 )† B6;FVIII / (B6;129S4-F8tm2Kaz) B6.FVIII / (B6.129S4-F8tm2Kaz) ‡ 129S1/SvmJ BALB/cAnCrl BALB/cOlaHsd BALB/cAnNTac BALB/cJ C57BL/6JCrl C57BL/6NCrl C57BL/6JBomTac C57BL/6JOlaHsd
/
/
129 BALB/c
C57BL/6
tm2Kaz
Breeder
N*
Baxter Innovations GmbH, Vienna, Austria Baxter Innovations GmbH, Vienna, Austria Baxter Innovations GmbH, Vienna, Austria Charles River Laboratories, Sulzfeld, Germany Charles River Laboratories, Sulzfeld, Germany Harlan Laboratories, San Pietro al Natisone, Italy Taconic Europe, Ejby, Denmark The Jackson Laboratory, Bar Harbor, ME, USA Charles River Laboratories, Sulzfeld, Germany Charles River Laboratories, Sulzfeld, Germany Taconic Europe, Ejby, Denmark Harlan Laboratories, San Pietro al Natisone, Italy
17 36 16 20 20 20 20 20 20 20 20 20
(11 M/6 F) (18 M/18 F) (8 M/8 F) (10 M/10 F) (10 M/10 F) (10 M/10 F) (10 M/10 F) (10 M/10 F) (10 M/10 F) (10 M/10 F) (10 M/10 F) (10 M/10 F)
F, female; M, male. *All with a body weight of 18–25 g, and age of 5–11 weeks. †Backcrossed with MAX-BAX speed congenics technology in cooperation with Charles River; full BALB/c background. ‡Backcrossed for eight generations; 98% C57BL/6 background.
Wild-type mice
Blood loss (mg)
1500
Hemophilic mice
1000 500
B6.FVIII–/–
B6;FVIII–/–
C.FVIII–/–
C57BL/6JOlaHsd
C57BL/6JBomTac
C57BL/6 N Crl
C57BL/6JCrl
BALB/cJ
BALB/c AnNTac
BALB/c OlaHsd
BALB/c AnCrl
129S1/SvmJ
0
Fig. 1. Blood loss in the tail-tip bleeding model in wild-type and FVIII / mice. 129S1/SvmJ and BALB/c mice showed almost no blood loss or interanimal variability in the tail-tip bleeding model, and there were no significant differences between substrains (P ≥ 0.146). The same was true for C57BL/6JBomTac and, to a certain extent, C57BL/6NCrl mice. C57BL/6JCrl and C57BL/6JOlaHsd mice showed a clearly higher total blood loss that was almost the same as that of hemophilic mice, with significantly higher interanimal variability than in C57BL/6JBomTac mice (P < 0.0001). Blood loss was lower in FVIII / mice on a BALB/c background (C.FVIII / ) than in FVIII / mice on a mixed background (B6; FVIII / ) or pure C57BL/6 background (B6.FVIII / ), and interindividual variation in B6.FVIII / mice was substantially lower than in B6;FVIII / mice (P < 0.0001). Furthermore, differences were observed between males (●) and females (○) in C57BL/6 and FVIII / mice.
The observed differences are most likely attributable to genetic diversity, differences in physiologic parameters not directly linked to coagulation (e.g. age), and environmental factors. A recently published example of the impact of genetic diversity is a spontaneous mutation in C57BL/ 6JOlaHsd mice (deletion of multimerin 1 and a-synuclein) that causes thrombus instability and impaired platelet function [13]. This finding is in line with the high blood loss observed in our tail-tip bleeding assay. Primary differences and changes with age have been described for many © 2014 International Society on Thrombosis and Haemostasis
physiologic parameters [14,15]. For example, platelet counts in male C57BL/6J mice increased from 1156 platelets lL-1 at 6 months of age to 2133 platelets lL-1 at 12 months of age, whereas those in female C57BL/6J mice remained stable from 6 to 12 months of age (1268 to 1229 platelets lL-1); other strains (e.g. 129S1/svInJ and BALB/cByJ) showed no major changes in platelet count with age [16]. Comparison of blood loss in our B6;FVIII / mice on an ill-defined C57BL/6 and 129S4 genetic background with that in B6.FVIII / mice backcrossed on a pure C57BL/6NCrl background demonstrated a marked reduction in interanimal variability (P < 0.0001; Fig. 1) and thus in the coefficient of variation (0.54% vs. 0.09%), as well as reductions in the bootstrap median and 95% confidence interval of variance in blood loss (353.43 mg and 294.58–40.89 mg vs. 70.70 mg and 28.49–93.27 mg, respectively). On the basis of these and the above-mentioned findings, we conclude that aligning the mouse strain and/or backcrossing of knockout mice with the preclinical model used to test drug efficacy could reduce data variability and thus the number of animals required per group while maintaining the power of statistical evaluation. We therefore consider it worthwhile to identify and use the mouse strain with the lowest variation and sex differences in the experimental models employed. The results of our study also stress the importance of stating the full nomenclature and the source of animals used in preclinical studies and scientific publications. Addendum A. Schiviz and W. Hoellriegl designed the protocol. A. Schiviz and P. Leidenmuehler performed the animal experiments. M. Schuster provided transgenic animals and data on the respective genetic background. A. Schiviz, M. Schuster, P. Leidenmuehler, and W. Hoellriegl collected, analyzed and interpreted data. D. Magirr performed statistical analysis. E. M. Muchitsch and
1942 A. Schiviz et al
W. Hoellriegl supervised research and interpreted data. A. Schiviz wrote the manuscript. All authors reviewed the manuscript for scientific content.
Disclosure of Conflict of Interests W. Hollriegl, P. Leidenmuhler, M. Schuster, E. M. Muchitsch, and A. Schiviz are full-time employees of Baxter Innovations GmbH. D. Magirr was contracted by Baxter to perform statistical analysis on the presented data. M. Schuster and E. M. Muchitsch hold patents on mouse models.
References 1 Parker ET, Lollar P. A quantitative measure of the efficacy of factor VIII in hemophilia A mice. Thromb Haemost 2003; 89: 480–5. 2 Tranholm M, Kristensen K, Kristensen AT, Pyke C, Rojkjaer R, Persson E. Improved hemostasis with superactive analogs of factor VIIa in a mouse model of hemophilia A. Blood 2003; 102: 3615–20. 3 Jesmok G, Cui ZH, Canivel D, Lollar P, Parker ET, Landskroner KA. Comparison of human rFVIII and murine rFVIII in a standardized FVIII dependent bleed model in FVIII–/– mice. J Thromb Haemost 2007; 5(Suppl. 2): P-M-030. 4 Greene TK, Schiviz A, Hoellriegl W, Poncz M, Muchitsch EM. Towards a standardization of the murine tail bleeding model. J Thromb Haemost 2010; 8: 2820–2. 5 Muchitsch EM, Schiviz A, Resch M, Hoellriegl W. Towards a Standardization of the Murine Tail Bleeding Model. 56th Annual Meeting of the Scientific and Standardization Committee of the ISTH, Cairo, Eygpt.2010.
6 Broze GJ Jr, Yin ZF, Lasky N. A tail vein bleeding time model and delayed bleeding in hemophiliac mice. Thromb Haemost 2001; 85: 747–8. 7 Ryan MJ, Didion SP, Davis DR, Faraci FM, Sigmund CD. Endothelial dysfunction and blood pressure variability in selected inbred mouse strains. Arterioscler Thromb Vasc Biol 2002; 22: 42–8. 8 Zurita E, Chagoyen M, Cantero M, Alonso R, Gonzalez-Neira A, Lopez-Jimenez A, Lopez-Jiminez JA, Landel CP, Benitez J, Pazos F, Montoliu L. Genetic polymorphisms among C57BL/6 mouse inbred strains. Transgenic Res 2011; 20: 481–9. 9 Hoover-Plow J, Shchurin A, Hart E, Sha J, Hill AE, Singer JB, Nadeau JH. Genetic background determines response to hemostasis and thrombosis. BMC Blood Disord 2006; 6: 6. 10 Ohkura N, Oishi K, Sakata T, Kadota K, Kasamatsu M, Fukushima N, Kurata A, Tamai Y, Shirai H, Atsumi G, Ishida N, Matsuda J, Horie S. Circadian variations in coagulation and fibrinolytic factors among four different strains of mice. Chronobiol Int 2007; 24: 651–69. 11 Rawle FE, Shi CX, Brown B, McKinven A, Tinlin S, Graham FL, Hough C, Lillicrap D. Heterogeneity of the immune response to adenovirus-mediated factor VIII gene therapy in different inbred hemophilic mouse strains. J Gene Med 2004; 6: 1358–68. 12 Bi L, Lawler AM, Antonarakis SE, High KA, Gearhart JD, Kazazian HH Jr. Targeted disruption of the mouse factor VIII gene produces a model of haemophilia A. Nat Genet 1995; 10: 119–21. 13 Tasneem S, Reheman A, Heyu N, Hayward C. C57BL/6OlaHsd mice with tandem deletion of the multimerin 1 and alpha-synoclein have impaired platelet function in vivo and in vitro that can be corrected by multimerin 1. Blood 2008; 112: 3926. 14 Mattson DL. Comparison of arterial blood pressure in different strains of mice. Am J Hypertens 2001; 14: 405–8. 15 The Jackson Laboratory. Mouse Phenome Database 2012. Available from URL: http://phenome.jax.org/. 16 Peters LL. Aging study: blood hematology in 30 inbred strains of mice: MPD:Peters4. The Jackson Laboratory 2007. Available from URL: http://phenome.jax.org/db/q?rtn=projects/details& sym=Peters4.
© 2014 International Society on Thrombosis and Haemostasis
Copyright of Journal of Thrombosis & Haemostasis is the property of Wiley-Blackwell and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use.
DOI: 10.1111/jth.12700
RECOMMENDATIONS AND GUIDELINES
Influence of genetic background on bleeding phenotype in the tail-tip bleeding model and recommendations for standardization: communication from the SSC of the ISTH € LER,* M. SCHUSTER,* E.-M. MUCHITSCH* and A . S C H I V I Z , * D . M A G I R R , † P . L E I D E N M UH ¨ W. HOLLRIEGL,* FOR THE SUBCOMMITTEE ON ANIMAL MODELS *Baxter Innovations GmbH; and †Medical Statistics Section, Center for Medical Statistics, Informatics and Intelligent Systems, Medical University of Vienna, Vienna, Austria
€ llriegl W, for the Subcommittee on Animal Models. To cite this article: Schiviz A, Magirr D, Leidenm€ uhler P, Schuster M, Muchitsch EM, Ho Influence of genetic background on bleeding phenotype in the tail-tip bleeding model and recommendations for standardization: communication from the SSC of the ISTH. J Thromb Haemost 2014; 12:1940–2.
The murine tail-tip bleeding model is widely used to measure bleeding time, blood loss, and survival after treatment [1–4]. Because of the considerable variation in data from these studies and thus the need for large sample sizes per group, and the lack of comparability of results from different laboratories, research groups have tried to standardize the model. In 2010, the SSC of the ISTH proposed standardized methods for determining tail bleeding time in mice. The SSC report standardized the tail clipping by endorsing the use of a warming chamber to evenly warm the anesthetized mouse, and assessing the influence of anesthesia and systolic blood pressure on blood loss [5]. We sought to test the ability of the standardized method to detect differences in bleeding phenotypes of wild-type mouse strains, which may show different bleeding phenotypes [6]. The relevance of the genetic background of inbred mouse strains or transgenic mice on phenotype has been described in various research fields. Mice may differ in physiologic parameters [7] or in response to triggers. Polymorphisms exist for substrains of the same inbred strains [8]. Genetic background also influences pathways that regulate thrombosis and hemostasis: differences were observed in global bleeding and thrombosis assays [9], coagulation and fibrinolytic factors [10], and susceptibility to treatment [11].
Correspondence: Eva-Maria Muchitsch, Baxter Innovations GmbH, Vienna DC Tower, Donau-City-Straße 7, 1220 Vienna, Austria. Tel.: +43 1 20100 247 2626; fax: +43 1 20100 557. E-mail: [email protected] Received 1 October 2013 Manuscript handled by: S. Eichinger Final decision: P. H. Reitsma, 28 July 2014
To investigate the ability of the tail-tip bleeding assay to detect the influence of genetic background on bleeding phenotype, we used hemophilic mice on different backgrounds, i.e. coagulation factor VIII (exon 17) knockout (FVIII / ) [12] mice on a BALB/c, C57BL/6, or mixed background, along with different substrains of wild-type C57BL/6 or BALB/c mice (Table 1). The mice were anesthetized with 100 mg kg-1 ketamine and 10 mg kg-1 xylazine and placed in a warming chamber. Two millimeters of the tail tip were clipped with a guillotine [4], the tails were immersed in warm saline (2 mL, 35 2 °C), and blood was collected over a period of 60 min; anesthesia was maintained as required by subcutaneous supplementation with one-third of the starting dose. Blood loss was determined gravimetrically. At the end of this observation period, the mice were removed from the chamber and humanely killed by cervical dislocation. Pairwise tests of equality of variance were performed with a bootstrap approach; the level of statistical significance was set to P = 0.05. C.FVIII / mice showed markedly lower median blood loss (365.7 mg) than B6.FVIII / (965.2 mg) and B6; FVIII / (890.4 mg) mice. For all three strains, median blood loss was lower in females than in males, with the largest sex difference (634 mg vs. 954 mg for females vs. males) and the highest interindividual variation being observed in B6;FVIII / mice (Fig. 1). In 129S1/SvmJ mice and the four substrains of BALB/c mice, median blood loss (23.5 mg and 10.0–19.0 mg, respectively) and interanimal variability were low. In contrast, median blood loss in C57BL/6 substrains varied from 36.0 mg (C57BL/6BomTac) to 469.5 mg (C57BL/6JCrl). Both the intersubstrain and interindividual ranges of blood loss (Fig. 1) and sex differences in median blood loss were high in C57BL/6 mice (e.g. 228 mg vs. 870 mg in C57BL/ 6JCrl and 76 mg vs. 765 mg in C57BL/6OlaHsd for females vs. males). © 2014 International Society on Thrombosis and Haemostasis
Mouse strains in the tail-tip bleeding model 1941 Table 1 Mouse strains Strain FVIII
Substrain C.FVIII (C.129S4-F8 )† B6;FVIII / (B6;129S4-F8tm2Kaz) B6.FVIII / (B6.129S4-F8tm2Kaz) ‡ 129S1/SvmJ BALB/cAnCrl BALB/cOlaHsd BALB/cAnNTac BALB/cJ C57BL/6JCrl C57BL/6NCrl C57BL/6JBomTac C57BL/6JOlaHsd
/
/
129 BALB/c
C57BL/6
tm2Kaz
Breeder
N*
Baxter Innovations GmbH, Vienna, Austria Baxter Innovations GmbH, Vienna, Austria Baxter Innovations GmbH, Vienna, Austria Charles River Laboratories, Sulzfeld, Germany Charles River Laboratories, Sulzfeld, Germany Harlan Laboratories, San Pietro al Natisone, Italy Taconic Europe, Ejby, Denmark The Jackson Laboratory, Bar Harbor, ME, USA Charles River Laboratories, Sulzfeld, Germany Charles River Laboratories, Sulzfeld, Germany Taconic Europe, Ejby, Denmark Harlan Laboratories, San Pietro al Natisone, Italy
17 36 16 20 20 20 20 20 20 20 20 20
(11 M/6 F) (18 M/18 F) (8 M/8 F) (10 M/10 F) (10 M/10 F) (10 M/10 F) (10 M/10 F) (10 M/10 F) (10 M/10 F) (10 M/10 F) (10 M/10 F) (10 M/10 F)
F, female; M, male. *All with a body weight of 18–25 g, and age of 5–11 weeks. †Backcrossed with MAX-BAX speed congenics technology in cooperation with Charles River; full BALB/c background. ‡Backcrossed for eight generations; 98% C57BL/6 background.
Wild-type mice
Blood loss (mg)
1500
Hemophilic mice
1000 500
B6.FVIII–/–
B6;FVIII–/–
C.FVIII–/–
C57BL/6JOlaHsd
C57BL/6JBomTac
C57BL/6 N Crl
C57BL/6JCrl
BALB/cJ
BALB/c AnNTac
BALB/c OlaHsd
BALB/c AnCrl
129S1/SvmJ
0
Fig. 1. Blood loss in the tail-tip bleeding model in wild-type and FVIII / mice. 129S1/SvmJ and BALB/c mice showed almost no blood loss or interanimal variability in the tail-tip bleeding model, and there were no significant differences between substrains (P ≥ 0.146). The same was true for C57BL/6JBomTac and, to a certain extent, C57BL/6NCrl mice. C57BL/6JCrl and C57BL/6JOlaHsd mice showed a clearly higher total blood loss that was almost the same as that of hemophilic mice, with significantly higher interanimal variability than in C57BL/6JBomTac mice (P < 0.0001). Blood loss was lower in FVIII / mice on a BALB/c background (C.FVIII / ) than in FVIII / mice on a mixed background (B6; FVIII / ) or pure C57BL/6 background (B6.FVIII / ), and interindividual variation in B6.FVIII / mice was substantially lower than in B6;FVIII / mice (P < 0.0001). Furthermore, differences were observed between males (●) and females (○) in C57BL/6 and FVIII / mice.
The observed differences are most likely attributable to genetic diversity, differences in physiologic parameters not directly linked to coagulation (e.g. age), and environmental factors. A recently published example of the impact of genetic diversity is a spontaneous mutation in C57BL/ 6JOlaHsd mice (deletion of multimerin 1 and a-synuclein) that causes thrombus instability and impaired platelet function [13]. This finding is in line with the high blood loss observed in our tail-tip bleeding assay. Primary differences and changes with age have been described for many © 2014 International Society on Thrombosis and Haemostasis
physiologic parameters [14,15]. For example, platelet counts in male C57BL/6J mice increased from 1156 platelets lL-1 at 6 months of age to 2133 platelets lL-1 at 12 months of age, whereas those in female C57BL/6J mice remained stable from 6 to 12 months of age (1268 to 1229 platelets lL-1); other strains (e.g. 129S1/svInJ and BALB/cByJ) showed no major changes in platelet count with age [16]. Comparison of blood loss in our B6;FVIII / mice on an ill-defined C57BL/6 and 129S4 genetic background with that in B6.FVIII / mice backcrossed on a pure C57BL/6NCrl background demonstrated a marked reduction in interanimal variability (P < 0.0001; Fig. 1) and thus in the coefficient of variation (0.54% vs. 0.09%), as well as reductions in the bootstrap median and 95% confidence interval of variance in blood loss (353.43 mg and 294.58–40.89 mg vs. 70.70 mg and 28.49–93.27 mg, respectively). On the basis of these and the above-mentioned findings, we conclude that aligning the mouse strain and/or backcrossing of knockout mice with the preclinical model used to test drug efficacy could reduce data variability and thus the number of animals required per group while maintaining the power of statistical evaluation. We therefore consider it worthwhile to identify and use the mouse strain with the lowest variation and sex differences in the experimental models employed. The results of our study also stress the importance of stating the full nomenclature and the source of animals used in preclinical studies and scientific publications. Addendum A. Schiviz and W. Hoellriegl designed the protocol. A. Schiviz and P. Leidenmuehler performed the animal experiments. M. Schuster provided transgenic animals and data on the respective genetic background. A. Schiviz, M. Schuster, P. Leidenmuehler, and W. Hoellriegl collected, analyzed and interpreted data. D. Magirr performed statistical analysis. E. M. Muchitsch and
1942 A. Schiviz et al
W. Hoellriegl supervised research and interpreted data. A. Schiviz wrote the manuscript. All authors reviewed the manuscript for scientific content.
Disclosure of Conflict of Interests W. Hollriegl, P. Leidenmuhler, M. Schuster, E. M. Muchitsch, and A. Schiviz are full-time employees of Baxter Innovations GmbH. D. Magirr was contracted by Baxter to perform statistical analysis on the presented data. M. Schuster and E. M. Muchitsch hold patents on mouse models.
References 1 Parker ET, Lollar P. A quantitative measure of the efficacy of factor VIII in hemophilia A mice. Thromb Haemost 2003; 89: 480–5. 2 Tranholm M, Kristensen K, Kristensen AT, Pyke C, Rojkjaer R, Persson E. Improved hemostasis with superactive analogs of factor VIIa in a mouse model of hemophilia A. Blood 2003; 102: 3615–20. 3 Jesmok G, Cui ZH, Canivel D, Lollar P, Parker ET, Landskroner KA. Comparison of human rFVIII and murine rFVIII in a standardized FVIII dependent bleed model in FVIII–/– mice. J Thromb Haemost 2007; 5(Suppl. 2): P-M-030. 4 Greene TK, Schiviz A, Hoellriegl W, Poncz M, Muchitsch EM. Towards a standardization of the murine tail bleeding model. J Thromb Haemost 2010; 8: 2820–2. 5 Muchitsch EM, Schiviz A, Resch M, Hoellriegl W. Towards a Standardization of the Murine Tail Bleeding Model. 56th Annual Meeting of the Scientific and Standardization Committee of the ISTH, Cairo, Eygpt.2010.
6 Broze GJ Jr, Yin ZF, Lasky N. A tail vein bleeding time model and delayed bleeding in hemophiliac mice. Thromb Haemost 2001; 85: 747–8. 7 Ryan MJ, Didion SP, Davis DR, Faraci FM, Sigmund CD. Endothelial dysfunction and blood pressure variability in selected inbred mouse strains. Arterioscler Thromb Vasc Biol 2002; 22: 42–8. 8 Zurita E, Chagoyen M, Cantero M, Alonso R, Gonzalez-Neira A, Lopez-Jimenez A, Lopez-Jiminez JA, Landel CP, Benitez J, Pazos F, Montoliu L. Genetic polymorphisms among C57BL/6 mouse inbred strains. Transgenic Res 2011; 20: 481–9. 9 Hoover-Plow J, Shchurin A, Hart E, Sha J, Hill AE, Singer JB, Nadeau JH. Genetic background determines response to hemostasis and thrombosis. BMC Blood Disord 2006; 6: 6. 10 Ohkura N, Oishi K, Sakata T, Kadota K, Kasamatsu M, Fukushima N, Kurata A, Tamai Y, Shirai H, Atsumi G, Ishida N, Matsuda J, Horie S. Circadian variations in coagulation and fibrinolytic factors among four different strains of mice. Chronobiol Int 2007; 24: 651–69. 11 Rawle FE, Shi CX, Brown B, McKinven A, Tinlin S, Graham FL, Hough C, Lillicrap D. Heterogeneity of the immune response to adenovirus-mediated factor VIII gene therapy in different inbred hemophilic mouse strains. J Gene Med 2004; 6: 1358–68. 12 Bi L, Lawler AM, Antonarakis SE, High KA, Gearhart JD, Kazazian HH Jr. Targeted disruption of the mouse factor VIII gene produces a model of haemophilia A. Nat Genet 1995; 10: 119–21. 13 Tasneem S, Reheman A, Heyu N, Hayward C. C57BL/6OlaHsd mice with tandem deletion of the multimerin 1 and alpha-synoclein have impaired platelet function in vivo and in vitro that can be corrected by multimerin 1. Blood 2008; 112: 3926. 14 Mattson DL. Comparison of arterial blood pressure in different strains of mice. Am J Hypertens 2001; 14: 405–8. 15 The Jackson Laboratory. Mouse Phenome Database 2012. Available from URL: http://phenome.jax.org/. 16 Peters LL. Aging study: blood hematology in 30 inbred strains of mice: MPD:Peters4. The Jackson Laboratory 2007. Available from URL: http://phenome.jax.org/db/q?rtn=projects/details& sym=Peters4.
© 2014 International Society on Thrombosis and Haemostasis
Copyright of Journal of Thrombosis & Haemostasis is the property of Wiley-Blackwell and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use.
