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Optimization of the Cuff Technique for Murine Heart Transplantation
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Summary June 26th, 2020
We introduce an inner tube approach to the cuff technique for mouse cervical heterotopic heart transplantation to help evert the vessel over the cuff. We found that cooperation between two experienced surgeons markedly shortens the operation time.
Transcript
Hello, everyone. My name is Dr.Zhongquan Qi.It's widely known that the cuff technique has been used as cardiac transplantation since in 1991. Although there have been great advancements in medical surgery and the new surgical technique, there are still some difficulties based on cuff technique.
It's a form of unveiling, where they will show you an optimotic half technique, which can significantly equal the surgical efficiency. The main procedures is inserting a temporarily in that tube into the visor. Proof read the ISO almost half.
Inserting the tube technique. A skilled surgeon can perform the operation within 35 minutes with 95%success rate. Then they can employ the Thermal Surgeon Operation Module, which further improves the efficiency of the operation showing in the skeptimic and the operation time will be shortened to about 35 minutes.
Thank you. And that's that ties the recipe and miles with pentobarbital. Use atraumatic mechanical clippers to remove the hair at the right later cervical region.
Use a sterile cotton tip applicator to wipe the surgical area with iodine antiseptic followed by 70%ethanol. Place the mouse in the supine position on the operation platform. Cover the mouse with sterile gauze.
Use an ophthalmic scissor to make a transverse incision from the lower one third NACMEC line to the right shoulder clavicle joint. Isolate the right external jungular vein with micro curved forceps to expose enough length. Cut off the brunches via electro coagulation.
And locate the vessel at the disto and using a six 2 0 six suture. Clamp the external junglar vein proximally using a powder clamp. And then translate the vein proximately to the ligature using a micro scissor.
Wash the vessel lumen with 100 units per milliliter 02 four Celsius degree heparin salient to remove any residual blood. Pull the external junglar vein through the vena cava using micro straight forceps. Insert the vein inner tube into the lumen as a stent and over the vessel wall over the curve with micro straight forceps.
Fix the irritant Faso endothelium at the proximal end of the curve using a circumferential eight to 06 suture. Use micro straight forceps to withdraw the vein inner tube from the vein vessel. Perform blunt dissection with micro forceps to isolate the right carotid artery, adjacent to the inner edge of the sternocleidomastoid.
Clamp the right carotid Artery proximally using a babcock clamp. Locate the carotid artery distally using a six to 06 suture. And use a micro scissor to transact the carotid artery proximally to the ligature.
Wash the carotid artery with 100 units per milliliter 02 four Celsius heparin salient to remove any residual blood. Pass the carotid artery through the artery curve and insert the artery inner tube into the artery vessel using micro straight forceps. Aim for the vessel over the curve using micro straight forceps.
Fix the inverted facile endothelium using a circumferential eight to 06 suture. Withdraw the artery inner tip from the artery vessel with micro straight forceps. Anaesthesia and its infection are the same as the recipient.
Make an abdominal mid line incision with an ophthalmic scissor and expose the abdominal cavity. Use micro curve forceps to expose the inferior vena cava. And then intravenously inject 200 microliter of 100 units per milliliter 02 four Celsius degree heparin salient per 20 gram of body weight through the inferior vena cava.
Perform thoracotomy with ophthalmic scissors. Cut off the ribs through the palettes Romita axillary light incisions. Flav the anterior chest wall outwards to expose the thoracic cavity.
Excise the thymus with micro forceps. Expose the aorta and then profuse 200 microliter of 100 units per milliliter 02 four Celsius degree heparin salient to the coronary artery through the aortic arch. Use a micro scissor to transect the ascending aorta at the beginning of the aortic arch.
Transect the pulmonary artery at the beginning of the two main branches with a micro scissor. Locate the superior vena cava and inferior vena cava approximately using a six to 02 suture and use a micro scissor to transect the vein distillate to the ligature. Locate the pulmonary veins together circumferentially using a single six, to 06 suture and cut off the vein branches distally to the ligature using a micro scissor.
Remove the heart graft from the surrounding soft tissues. Place the donor heart upside down into the right neck region of the recipient. Input the pulmonary artery of the donor heart to a six to 06 loop with micro straight forceps.
Wrap the vessel lumen around the vena cava and then tighten the six to 06 suture loop around the calf to bend the vessel joint. Perform anastomosis of the aorta of the graft and artery curve. Release the clumped jugular vein followed by clumped jugular artery.
Keep the vessel joined and twisted and ensure that the blood flow is unobstructed. Set the passing heart graft into the subcutaneous space and then suture the incision. Record the time to normal sinus rhythm and the preservation of normal sinus rhythm for at least five minutes after clamp release to monitor the post operative graft function.
Place the recipient alone on the one blanket until the recipient wakes up from anesthesia. Taken together, although our surgery uses cuff techniques that have been reported, we established the inner core technique based on this. Reduce the difficulty in surgery and shorten the anastomosis time by optimization technique.
We suggested the double surgery incorporation motto to further improve the efficiency of the operation shown in the schematic. And the operation time will be shortened to about 25 minutes. A fully MHC mismatched BALBC allograft heart can be rejected with eight days after transplantation into C57BL6 recipient mice.
Syngeneic heart transplants used in our research survived more than 100 days except one rare case due to a 15%weight loss compared to normal weight before the operation. Histopathology of the allograft on day seven revealed typical features of acute rejection, including a heavy cellular infiltrate with tissue necrosis. Syngeneic grafts are near normal with no evidence of myocyte necrosis or inflammatory cell infiltration.
100 days after syngenetic heart transplant, the vascular endothelium of anastomosis site can be collected and stained by immunofluorescent. In this analysis, no averse snaring of vascular wall, thrombosis or thickening of the intima were observed. Electro microscopic imaging revealed that a smooth endothelium and a regular longitudinal crust formation with the endothelial cells arranged neatly and closely with no obvious sediment on the surface.
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