Figure 1: Experimental set up for isolation of murine HSC. (A) Prior starting, the work space is prepared and the sterilized instrumentation that is necessary for the surgery placed in clear order. The fur of anesthetized mouse is shaved, the mouse placed onto the working area and the surgery area covered with fluid-impermeable, self-adhesive drapes. (B) The abdomen is opened with a midline laparotomy and (C) the abdominal skin lifted with forceps and removed with a sterile scissors. (D) The liver is uncovered and (E) the ventral side of the liver is lifted with a humidified cotton swab so that the portal vein is freely exposed for execution of the perfusion.
Figure 2: Liver perfusion. (A) A canula is carefully plugged into the portal vein without rupturing the vessel wall and (B) the canula is positioned into a stable position by fixing it with tweezers. (C) The inferior vena is cut with scissors and (D) the peristaltic pump started allowing the perfusion buffer to flow freely through the liver. During this process, the organs are fixed with tweezers. (E) The perfusion is continued until the liver is cleaned from blood that is visible by a change from dark red to brown. (F) During the complete liver perfusion, the perfusion buffers are pre-warmed to 37*C. This is done by storing them in a heated water bath.…show more content… (A) After successful digestion, the liver has a very light colour. (B) The canula is removed and the liver carefully uncovered taking special care not to rupture the esophagus. (C) The liver is grabbed with surgical tweezers and removed from the abdominal cavity by cutting the ligaments with small scissors. (D) The liver is then placed into a petri dish with some buffer and (E) the Glisson’s capsule removed to release the dispersed cells. (F) The gall bladder is carefully removed and (G) the digested liver grabbed with a scissor and intensively shaked to release the
