Name: Inferior Vena Cava (IVC) Ligation for Venous Thrombosis Analysis in Mouse
Uploaded: 2024-01-05T13:30:00
Description: Watch this Scientific Journal Video about Inferior Vena Cava IVC Ligation for Venous Thrombosis Analysis in Mouse at JoVE.com

inferior vena cava (ivc) ligation for venous thrombosis analysis in mouse

Optimalisert venøs tromboseanalyse i en musekreftmodell ved bruk av vaskulære klips

Cancer-associated venous thromboembolism (VTE) 
Venous thromboembolism (VTE) risk is 4 to 7 times higher in cancer survivors compared to the general population1,2,3. This condition proves fatal in one out of seven patients with cancer. The incidence of VTE varies depending on the type of cancer and the tumor burden and is highest among patients with pancreatic and gastric cancers4.
Cancer-associated VTE in cancer patients has prognostic significance. It is associated with unfavorable overall survival in the first year after a cancer diagnosis, even after adjusting for age, race, and stage of underlying cancer5. These findings highlight the importance of examining cancer associated VTE and the need to probe its mechanism using an animal model. The translational relevance of this area is further emphasized by the fact that VTE in cancer patients is preventable and treatable with thromboprophylaxis and antithrombotic therapy6.
Animal models of cancer and venous thrombosis 
Cancer models are conventionally termed xenografts, which entail the injection of cancer cells in mice. The injection of cancer cells at a site like its origin is referred to as an orthotopic model, while at a different site (subcutaneous plane over the flank) is known as a heterotopic model. The species of origin of cancer cells determines them as an allogeneic model, such as the HT-29 cell line (human colon cancer)7,8,9. On the contrary, syngeneic models use the murine cancer cell lines, including RenCa and MC-38 cell lines3,10.
The literature has described arterial, venous, and capillary thrombosis models in rodents. Venous thrombosis is induced in the inferior vena cava (IVC) by mechanical injury (guide wire) or complete IVC ligation, chemical (Ferric chloride), or electrolytic injury. Ferric chloride-induced thrombosis or IVC ligation represents complete occlusion models. The latter results in the stasis of blood and inflammatory infiltrates in veins11,12,13. The complete ligation model results in a high rate of thrombosis formation in 95% to 100% of mice. The partial IVC ligation model might include interruption of lateral iliolumbar branches, and the venous return is abrogated by applying suture ligations in the distal target points of IVC12. Sometimes, a space holder is used to interrupt the venous return partially. However, the thrombus weight is inconsistent in the current partial occlusion model, resulting in high variability in clot weights and heights12,14.
Both these large vein mechanical models (partial and complete) have limitations. First, IVC ligation (stasis model) often results in hypotension. The blood gets shunted through vertebral veins. Though in experienced hands, the mortality with this model ranges from 5%-30%, with the higher rate expected during the learning curve. Importantly, the complete occlusion model does not reproduce deep vein thrombosis (DVT) in humans, where a thrombus typically is nonocclusive. Complete occlusion is likely to alter hemorheological factors and pharmacodynamic parameters, altering the bioavailability of compounds at the local site. Due to these limitations, complete occlusion models may not be optimal for testing novel chemical compounds for therapeutic purposes and drug discoveries12.
It should be noted that to provide a more clinically relevant murine model of venous thrombosis with decreased flow with endothelial damage, a venous thrombosis model has been introduced, where DVT is triggered by the restriction of blood flow in the absence of endothelial disruption. The model was validated by scanning electron microscopy15. A preferred clinically relevant thrombosis model is one with near complete thrombosis that enables drug discoveries. The clot formation in the current partial occlusion models is inconsistent, resulting in high variability in the clot weight and heights12,16. Furthermore, the clot weight is variable with the conventional methods, requiring&#160;more&#160;mice per studies12.
Previous cancer-associated thrombosis models focused on colon, pancreatic, and lung cancer and were all complete occlusion models17,18,19. This manuscript modifies the partial occlusion thrombosis model to provide clots with lower variability and mouse mortality (Figure 1). Former studies used allogeneic cancer cell lines on immunocompromised athymic mice background19,20,21. This manuscript uses an MC-38 cell syngeneic xenograft in C57Bl6/J mice, which allows the use of immunocompetent mice and examination of immune components to thrombogenesis.

Forfatter Spotlight: Fremme kreftassosiert tromboseforskning i gnagermodeller 

Venøs tromboseanalyse i en musemodell av kreft

This research aims to enhance the rodents model for venous thrombosis focusing on cancer-associated thrombosis. It introduces a refined surgical technique using vascular clips in a syngeneic colon cancer xenograft mouse model to improve consistency and reproducibility, advancing cancer-associated thrombosis mechanistic understanding and drug discovery. Although the flow reduction model was introduced in the IVT stenotic models in 2012, no significant progress has been made in cancer-associated thrombosis models in the last 10 years.This was the rationale for this endeavor. In the field of cancer-associated thrombosis research, current technologies include advanced imaging techniques, like scanning, electron microscopic images, molecular biology tools, such as immunofluorescent staining, precision surgical instrument, data analysis software and data interpretation, and animal welfare. The current experimental challenges include variability in clot weights, a high mortality rate, and a prolonged learning curve in rodent models of venous thrombosis.Additionally, the venous flow direction and clot formation mechanisms differ from clinical cases. Our research protocol bridges gaps in rodent models of cancer-associated thrombosis. It introduces a novel surgical technique using vascular clips in xenograft models, yielding more consistent and reproducible thrombosis outcomes.This enhances our understanding of cancer-related thrombosis and drug discovery in translational research.

Watch this Scientific Journal Video about Inferior Vena Cava IVC Ligation for Venous Thrombosis Analysis in Mouse at JoVE.com

Inferior Vena Cava (IVC) Ligation for Venous Thrombosis Analysis in Mouse  

Inferior vena cava (IVC) ligering for venøs tromboseanalyse hos mus

For å begynne, bruk en fin saks for å lage et snitt på bukhuden til en bedøvet mus, fra xiphoidal eminens til blæren. Med et par atraumatiske tang, klem bukhinnen halvveis langs hudsnittet, og sørg for et tilstrekkelig gap mellom snittet og de overliggende tarmene. Deretter åpner du bukhinnen i lengderetningen langs den avaskulære linea alba.Bruk deretter våte bomullstips for å trekke ut tarmene forsiktig. Deretter slipper 200 til 300 mikroliter normal saltvann over tarmene, og utforske ruten til urinlederne dårligere vena cava, eller IVC, aorta og nyreårer. Cauterize de bilaterale grenene distalt for sammenløpet av venstre nyrevene og infrarenal inferior vena cava.Sjekk urinlederne og vaskulaturen for eventuelle oser. For å forhindre potensielle komplikasjoner av spinal iskemi og claudicatio, la lumbale grener være intakte. Sjekk for sidegrenene, inkludert det bilaterale livmorhornet og hypogastriske kar.Deretter bruker du en atraumatisk tang med lukket spiss for forsiktig å passere en seks-null polypropylen sutur gjennom det avvaskulære planet to til tre ganger. Påfør forsiktig trykk med en bomullstupptamponade for å kontrollere eventuell osing. Til slutt, pass en fem-null polypropylen sutur gjennom flyet med forsiktig coddle til kraniale bevegelser for å utvide den.Plasser vaskulære klemmer over suturen for å sikre tilstrekkelig plass. Koagulasjonsvekter i den xenotransplanterte eksperimentgruppen viste en økning på 1,5 ganger i forhold til kontrollmusene. Immunfluorescenstester viste at IVC fra kontrollgruppen hadde intakt CD31 og fibrin som delvis opptok karlumen.CD31-uttrykket var ikke intakt i eksperimentgruppen, og veggen viste fibrinfarging. Kontrollgruppen hadde signifikant lavere fibrinuttrykk sammenlignet med eksperimentgruppen. Ultralyd grafisk analyse av musene viste ligert infrarenal vena cava av en kontrollgruppe mus hadde en delvis blodpropp.Den eksperimentelle gruppen hadde imidlertid en stor blodpropp som nesten okkluderte den infrarenale vena cava.

Research

JoVE Journal

Cancer Research

Inferior Vena Cava (IVC) Ligation for Venous Thrombosis Analysis in Mouse

Venous Thrombosis Assay in a Mouse Model of Cancer

This methodology paper highlights the surgical nuances of a rodent model of venous thrombosis, specifically in the context of cancer-associated thrombosis ...