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Updated: May 22, 2026

Isolation of Human Hepatocytes by a Two-step Collagenase Perfusion Procedure
Published on: September 3, 2013
A novel cannulation technique for isolation of human hepatocytes from explanted diseased whole livers
D C Kehr1, N Raschzok, I M Sauer
1General, Visceral, and Transplantation Surgery, Experimental Surgery and Regenerative Medicine, Charité-Campus Virchow, Universitätsmedizin Berlin, Berlin, Germany.
Abstract:
Diseased human organs explanted during liver transplantation can be used as a cell source for basic research and future therapeutic applications in regenerative medicine. Enzymatic digestion using the perfusion technique has become the gold standard in liver cell isolation. Usually the portal vein is used as a vascular access for liver cell isolation from explanted livers, that were rejected from whole organ transplantation. No special techniques are required for cannulation; the cannulas are simply introduced into the vessels and a ligature is then thrown around the vessel to secure the cannulation. This method is not applicable to organs explanted during liver transplant surgery, because as much of the vessels as possible has to be kept in situ, to facilitate anastomosis of the new organ. Therefore, when perfusing the explanted organ, normal perfusion catheters are easily displaced and a more complex "vascular reconstruction" must be performed to secure hold of the catheters. We established a novel cannulation technique using commercially available Foley catheters for liver cell isolation from diseased whole organs explanted during transplant surgery. We evaluated this technique in 15 diseased organs. 5 were isolated in the conventional setting and 10 were cannulated using Foley catheters. The average cannulation time was significantly shortened using Foley catheters compared with the conventional approach (12 ± 5.2 min vs 40 ± 14.1 min; P = .0001). Foley catheter cannulation is fast, simple, and efficient. It appears to be favorable for hepatocyte isolation from diseased whole livers or from explanted organs with technically difficult vascular access.
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