Orthotopic mouse liver transplantation to study liver biology and allograft tolerance

Abstract

Orthotopic liver transplantation in the mouse is a powerful research tool that has led to important mechanistic insights into the regulation of hepatic injury, liver immunopathology, and transplant tolerance. However, it is a technically demanding surgical procedure. Setup of the orthotopic liver transplantation model comprises three main stages: surgery on the donor mouse; back-table preparation of the liver graft; and transplant of the liver into the recipient mouse. In this protocol, we describe our procedure in stepwise detail to allow efficient completion of both the donor and recipient operations. The protocol can result in consistently high technical success rates when performed by personnel experienced in the protocol. The technique can be completed in ∼2-3 h when performed by an individual who is well practiced in performing mouse transplantation in accordance with this protocol. We have achieved a perioperative survival rate close to 100%.

Graft‐infiltrating PD‐L1hi cross‐dressed dendritic cells regulate antidonor T cell responses in mouse liver transplant tolerance

“…For all (liver transplant) experiments, age‐matched animals were distributed randomly into experimental groups. Liver transplants were performed in male B6 donors to male C3H recipients because for anatomic/surgical reasons, male B6 (or B10) into male C3H is the model of choice for orthotopic allogeneic liver transplantation in the mouse . After surgery, each mouse was housed alone for the first week.…”

“…Liver transplants were performed in male B6 donors to male C3H recipients because for anatomic/ surgical reasons, male B6 (or B10) into male C3H is the model of choice for orthotopic allogeneic liver transplantation in the mouse. (27,(32)(33)(34) After surgery, each mouse was housed alone for the first week. All surgeries were performed during the light cycle without fasting.…”

Acetyl‐3‐Aminoethyl Salicylate Ameliorates Hepatic Ischemia/Reperfusion Injury and Liver Graft Survival Through a High‐Mobility Group Box 1/Toll‐Like Receptor 4–Dependent Mechanism

“…Nonimmunosuppressed WT and TLR4 KO C57BL/6 mice were used to construct a murine orthotopic liver transplant model as recently described by Yokota et al, with minor modifications . Briefly, the transplant procedure consisted of 3 stages: Donor liver graft resection. Cold preservation. Recipient liver graft implantation …”

“…For recipient liver graft implantation, because portal vein clamping (PVC) beyond 20‐30 minutes during liver graft implantation can lead to recipient death, all PVC procedures were conducted within a strict 20‐minute time frame (mean PVC time, 18.1 minutes; range, 16.1‐20.0 minutes) . Then, ac3AESA‐treated recipient mice were administered IV ac3AESA (15 mg/kg) according to the following 2 protocols: the longterm ac3AESA protocol—30 minutes after transplant (day 0) followed by ac3AESA (15 mg/kg) daily until day 10 after transplant; or the short‐term ac3AESA protocol—30 minutes after transplant (day 0) followed by ac3AESA (15 mg/kg) daily until day 3 after transplant.…”

“…Recipient liver graft implantation. (26) Donor mice were anesthetized by sodium pentobarbital (50 mg/kg), and donor livers were resected and flushed with 30 mL of University of Wisconsin (UW) solution. Donor livers then underwent cold preservation (4°C) for 12, 18, or 24 hours in the UW solution prior to orthotopic transplantation.…”