These videos demonstrate the cannulation, wash (HBSS), digestion (collagenase/DMEM), excision, and purification steps described in the associated protocol. All procedures were performed in an IACUC-sanctioned lab and in complete accordance with IACUC regulations.
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Isolating Primary Mouse Hepatocytes
[1] Cannulation of portal vein and cutting of IVC; HBSS perfusion
[2] IVC clamp to confirm successful cannulation; still HBSS
[3] IVC clamp after ~1 minute of collagenase digestion
[4] IVC clamp after ~2 minutes of collagenase digestion
[5] IVC clamp after ~3 minutes of collagenase digestion
[6] IVC clamp after ~5.5 minutes of collagenase digestion
[7] Excision of liver immediately following end of digestion
[8] Tearing and shaking of excised liver in collagenase media
[9] Initial filtration and resuspension of liberated cells
[10] First round of washing and centrifugation
[11] Second round of washing and centrifugation
[12] Third (final) round of washing and centrifugation