Authors: Jae-Jin Cho¹; Brigid Joseph¹; Baljit Singh Sappal¹; Ranjit K. Giri¹; Richard Wang²; John W. Ludlow²; Mark E. Furth²; Robert Susick²; Sanjeev Gupta¹

Source: Liver International, Volume 24, Number 4, August 2004 , pp. 361-370(10)

Publisher: Wiley-Blackwell

Abstract:

Cho J-J, Joseph B, Sappal BS, Giri RK, Wang R, Ludlow JW, Furth ME, Susick R, Gupta S. Analysis of the functional integrity of cryopreserved human liver cells including xenografting in immunodeficient mice to address suitability for clinical applications.

Liver International 2004: 24: 361–370. © Blackwell Munksgaard 2004 Abstract: Background:

The availability of well-characterized human liver cell populations that can be frozen and thawed will be critical for cell therapy. We addressed whether human hepatocytes can recover after cryopreservation and engraft in immunodeficient mice. Methods:

We isolated cells from discarded human livers and studied the properties of cryopreserved cells. The viability of thawed cells was established with multiple in vitro assays, including analysis of liver gene expression, ureagenesis, cytochrome P450 activity, and growth factor-induced cell proliferation. The fate of transplanted cells was analysed in immunodeficient NOD-SCID mice. Results:

After thawing, the viability of human hepatocytes exceeded 60%. Cells attached to culture dishes, proliferated following growth factor stimulation and exhibited liver-specific functions. After transplantation in NOD-SCID mice, cells engrafted in the peritoneal cavity, a heterologous site, as well as the liver itself, retained hepatic function and proliferated in response to liver injury. Transplanted hepatocytes were integrated in the liver parenchyma. Occasionally, transplanted cells were integrated in bile ducts. Conclusions:

Cryopreserved human liver cell showed the ability to retain functional integrity and to reconstitute both hepatic and biliary lineages in mice. These studies offer suitable paradigms aimed at characterizing liver cells prior to transplantation in people.

Keywords: cryopreservation; hepatocyte; human; mouse; transplantation

Document Type: Research article

DOI: http://dx.doi.org/10.1111/j.1478-3231.2004.0938.x.

Affiliations: 1: Departments of Medicine and Pathology, Marion Bessin Liver Research Center, Albert Einstein College of Medicine, Bronx, NY, USA and 2: Vesta Therapeutics Inc. (formerly Incara Cell Technologies), Research Triangle Park, NC, USA

Publication date: 2004-08-01

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