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Summary

Abstract

Introduction

Protocol

Representative Results

Discussion

Acknowledgements

Materials

References

Medicine

Veno-Venous Extracorporeal Membrane Oxygenation in a Mouse

Published: October 24th, 2018

DOI:

10.3791/58146

1Department of Cardiothoracic, Transplantation, and Vascular Surgery, Hannover Medical School, 2Department of Pneumology, Hannover Medical School
* These authors contributed equally

Here we present a protocol describing the technique of veno-venous extracorporeal membrane oxygenation (ECMO) in a non-intubated, spontaneously breathing mouse. This murine model of ECMO can be effectively implemented in experimental studies of acute and end-stage lung diseases.

The use of extracorporeal membrane oxygenation (ECMO) has increased substantially in recent years. ECMO has become a reliable and effective therapy for acute as well as end-stage lung diseases. With the increase in clinical demand and prolonged use of ECMO, procedural optimization and prevention of multi-organ damage are of critical importance. The aim of this protocol is to present a detailed technique of veno-venous ECMO in a non-intubated, spontaneously breathing mouse. This protocol demonstrates the technical design of the ECMO and surgical steps. This murine ECMO model will facilitate the study of pathophysiology related to ECMO (e.g., inflammation,bleeding and thromboembolic events). Due to the abundance of genetically modified mice, the molecular mechanisms involved in ECMO-related complications can also be dissected.

Extracorporeal membrane oxygenation (ECMO) is a temporary life support system that takes over functions of the lungs and heart to allow adequate gas exchange and perfusion. Hill et al1 described the first use of ECMO in patients in 1972; however, it only became widely used after its successful application during the H1N1 influenza pandemic in 20092. Today, ECMO is routinely used as a lifesaving procedure in end-stage heart and lung diseases3. Veno-venous ECMO is increasingly employed as an alternative to invasive mechanical ventilation in awake, non-intubated, spontaneously breathing patients with....

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Experiments were performed on male C57BL/6 mice, aged 12 weeks. This study was conducted in compliance with guidelines of the German Animal Law under Protocol TSA 16/2250.

1. Materials Preparation

NOTE: All steps are performed under clean, non-sterile conditions. Sterile conditions would be required if animal is to be survived postoperatively.

  1. Introduce 3 fenestrations into a 2-Fr polyurethane tube using a surgical blade under a microscope with 16X .......

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This protocol describes the method of veno-venous ECMO in a mouse. This model is reliable and reproducible, and compared to our previously described model of CPB with respiratory and circulatory arrest12,13, it is less technically demanding to establish.

ECMO flow in the venous system was maintained between 1.5 and 5 mL/min. The mean arterial pressure was kept between 70.......

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Previously, we described a successful model of CPB in a mouse12,13. To implement such a model for acute or end-stage lung disorders we developed an easy-to-use veno-venous ECMO circuit for mice. Different to the CPB model, veno-venous ECMO does not require complicated surgical procedures such as sternotomy and clamping of the aorta, thus reducing the risk of wound bleeding in a fully heparinized animal. To avoid embolization of the oxygenator with blood clots, 2........

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This project was supported by KFO 311 Grant from Deutsche Forschungsgemeinschaft.

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Name Company Catalog Number Comments
Sterofundin B.Braun Petzold GmbH PZN:8609189 in 1:1 with Tetraspan
Tetraspan 6% Solution B. Braun Melsungen AG PZN: 05565416 in 1:1 with Sterofundin
Heparin Natrium 25.000 Ratiopharm GmbH PZN: 3029843 2,5 IU per ml of priming
NaHCO3 8,4% Solution B. Braun Melsungen AG PZN: 1579775 3% in priming solution
Carprofen Zoetis Inc., USA PZN:00289615 5mg/kg/BW
1 Fr PU Catheter Instechlabs INC., USA C10PU-MCA1301 carotide artery
2 Fr PU Catheter Instechlabs INC., USA C20PU-MJV1302 jugular vein
8-0 Silk suture braided Ashaway Line & Twine Co., USA 75290 ligature
Isoflurane Piramal Critical Care GmbH PZN:9714675 narcosis
Spring Scissors - 6mm Blades Fine Science Tools GmbH 15020-15 instruments
Spring Scissors - 2mm Blades Fine Science Tools GmbH 15000-03 instruments
Halsted-Mosquito Hemostat Fine Science Tools GmbH 13009-12 instruments
Dumont #55 Forceps Fine Science Tools GmbH 11295-51 instruments
Castroviejo Micro Needle Holder - 9cm Fine Science Tools GmbH 12060-02 instruments
Micro Serrefines Fine Science Tools GmbH 18555-01 instruments
Bulldog Serrefine Fine Science Tools GmbH 18050-28 instruments
Isoflurane Vaporizer Drager 19.1 Drägerwerk AG & Co. KGaA anesthesia 1,3 -2,5%
Multichannel Data Aquisition Device with ISOHEART Software Hugo Sachs Elektronik GmbH, Germany invasive pressure, ECG, t °C
i-STAT portable device Abbott Laboratories, Lake Bluff, Illinois, USA blood gas analysis
i-STAT CG4+ and CG8+ cartridges Abbott Laboratories, Lake Bluff, Illinois, USA blood gas analysis
C57Bl/6 mice, male, 30 g, 14 weeks old Charles River Laboratories housed 1 week before

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