Developing a Humanized Mouse Model with Human Liver Cells and Immune Cells

First, attach one end of a sterile extension tube to a 30-gauge needle, and the other end to a 1-milliliter syringe. Fill the syringe with the suspension of pooled HEPs and HSPCs. Fit the syringe in the notch of a repetitive dispensing pipette, and adjust the dispenser to dispense 10 microliters in each press. Shave each mouse as outlined in the text protocol, and scrub the left side of the body of each mouse with 10% povidone-iodine, followed by 70% isopropyl alcohol.

Using Vannas-type scissors, make a small incision in the skin, muscle, and peritoneum at the left of the peritoneal wall to enter the peritoneal cavity approximately 5 millimeters below the lower edge of the rib cage. Locate the spleen and use forceps to pull it slightly to the operating area for easy access, and insert the 30-gauge needle into the lower pole of the spleen.

Next, unlock the plunger of the dispensing pipette, and dispense 10 microliters of the volume at a time, with a limit of 60 to 80 microliters per spleen. Retract the needle slowly, and clip the spleen with ligating clips using a ligation applier. Then, use cotton-tipped applicators, wetted with sterile PBS, to push the spleen back into the body cavity. Use an interrupted suture pattern to close the muscle layer of the abdominal wall. Accomplish skin closure with an interrupted suture pattern using non-absorbable sutures.