Elsevier

Surgery

Volume 127, Issue 2, February 2000, Pages 193-199
Surgery

Original Communications
Multiple intrasplenic hepatocyte transplantations in the dalmatian dog*,**

https://doi.org/10.1067/msy.2000.102043Get rights and content

Abstract

Background: Hepatocyte transplantation is an attractive potential treatment for liver-based inborn errors of metabolism and for fulminant hepatic failure. Dalmatian dogs have a metabolic error that results in hyperuricosuria. This report focuses on the effect of multiple, sequential intrasplenic transplants of fresh and cryopreserved hepatocytes in dalmatians. Methods: Dalmatians underwent intrasplenic hepatocyte transplantation with hepatocytes taken from healthy mongrels. Dalmatian urinary uric acid excretion was measured preoperatively, and this served as the control value. Three hepatocyte transplantations were performed at 30-day intervals—the first with freshly isolated cells,and both the second and the third with cryopreserved hepatocytes from the same donor. Urinary uric acid excretion was measured postoperatively twice per week. Results: The urinary uric acid excretion decreased an average of 54% after the first hepatocyte transplantation. The effect was transient and lasted an average of 22 days (range, 19-50 days). Subsequent intrasplenic hepatocyte transplantation with cryopreserved hepatocytes resulted in similar decreases in urinary uric acid excretion. Each transplant resulted in a significant decrease in urinary uric acid excretion when compared with baseline values (P = <.001). Conclusions: Sequential intrasplenic hepatocyte transplantation is feasible in this model. This method provided a significant, but transient, correction in urinary uric acid excretion that was similar with either fresh or cryopreserved hepatocytes. A substantial biologic effect provided by cryopreserved hepatocytes has important implications in clinical hepatocyte transplantation. (Surgery 2000;127:193-9.)

Section snippets

Material and methods

Mongrel canines (n = 6) and non-littermate dalmatian dogs (n = 6) (Triple C Farms, St. Joseph, Ill) with an average body weight of 20 kg were maintained at the Biological Resource Laboratory of the University of Illinois at Chicago. Dogs were fed a standard amount of food (Teklad 25% Lab Dog Diet; Harlan Teklad, Madison, Wis) each morning. All procedures were approved by the University of Illinois Institutional Animal Care and Use Committee and conformed to the guidelines for humane care of

Results

After hepatocyte isolation, cell viability averaged 91% (± 5). The total number of hepatocytes recovered after isolation ranged from 32 to 60 × 109 cells. The plating efficiency of freshly isolated hepatocytes was estimated at greater than 80%. The total cell load administered during fresh HTx averaged 184 ± 16 (×106) cells/kg. Overall, CHx were 58 ± 14% viable after thawing. Consistently higher CHx cell viability (as high as 82%) was achieved near the end of the study, reflecting our steep

Discussion

The most limiting feature of hepatocyte transplantation in the dog is the inability to safely transplant a cell mass that is large enough to provide a prolonged therapeutic response. The dog possesses a highly reactive splanchnic vasculature when compared with human beings. The maximum safe number of cells that can be transplanted in a single setting has been determined by our previous work and is the basis for the method and number of hepatocytes used in this study.12 Since this number of

Acknowledgements

We thank Raymond Pollak, MB, and Jose Arruda, MD for their constructive comments and Jeff Fortman, DVM, for his expert help with the animals.

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    *

    Supported in part by the Eleanor B. Pillsbury Foundation.

    **

    Reprint requests: Enrico Benedetti, MD, Division of Transplantation, University of Illinois at Chicago, 840 S Wood St M/C 958, Chicago, IL 60612.

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