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Strategies of hepatocyte transplantation

100%

Ostrowska A.

Postępy Higieny i Medycyny Doświadczalnej

2000

tom 54

nr 2

149-181

Hepatic parenchymal cell transplantation is a promising method for managing patients with the acute liver failure and may create a bridge to whole organ grafting. Elimination or reduction of the immunogenicity of the hepatocytes would permit long-term graft survival without the neet of non-specific immunosuppression. The presented experimental evidence suggests that the modulation of hepatocyte immunogenicity by purification and cryopreservation reduces the allorespense to hepatocytes both in vitro and in vivo.Identification of inoculated cells facilitates long-term monitoring of their localization and metabolic activity.

Characterization of human hepatocytes isolated from non-transplantable livers

80%

Laba A. , Ostrowska A. , Patrzalek D. , Paradowski L. , Lange A.

Archivum Immunologiae et Therapiae Experimentalis

2005

tom 53

nr 5

442-453

The successful use of hepatocytes depends on a reliable demonstration of the functional and morphological integrity of isolated cells. Herein we investigated whether the isolation and cryopreservation of primary human hepatocytes can compromise cell viability and liver-specific characteristics. Hepatocytes were isolated from encapsulated human liver segments by a modified 2-step perfusion technique. Isolated cells were Percoll-purified, cryopreserved, and stored in liquid nitrogen for 1?12 months. For rapid assessment of fresh and cryopreserve/thawed hepatocyte yield and viability, the cells were stained with trypan blue or labeled with fluorochromes. For immunocytochemical analysis, the cells were labeled with monoclonal antibodies for the presence of the following antigens and chemokines: CD3, CD45Ro, CD45Ra, CD34, CD68, CD90, CD95, CD20, HLA-DR, Ki67, PCNA, Bcl-2, p53, CXCR3, CXCR4, and SDF-1. The cells were tested for several specific functions, such as ureagenesis, energy status, MTT activity, lactate dehydrogenase leakage, and total CYP450 content. Assessment of both freshly isolated (Percoll-purified) and cryopreserved/thawed hepatocytes revealed a low constitutive level of contamination by non-parenchymal cells compared with crude (unpurified) preparations and tissue sections. All viable hepatocytes showed intact morphology and retained CYP450 protein, energy status, and urea synthesis. Modifications in hepatocyte preparations, such as depletion of dead, damaged, and non-parenchymal cells, improves cell purity, which can be adapted to further evaluation of hepatocyte immunogenicity. These data illustrate the importance and feasibility of human hepatocyte banking.

Molecular mechanisms regulating transport of low molecular substances in hepatocyte

70%

Zwierz K. , Wielgat P. , Borzym-Kluczyk M.

Postępy Higieny i Medycyny Doświadczalnej

2003

tom 57

nr 1

91-116

Transporters of sinusoidal and lateral membranes mediate uptake of inorganic and organic substances to hepatocyte. In hepatocyte lipophilic compounds are biotransformed (hydroxylated and conjugated) and excreted into bile and urine. Vectorial movement of solutes and water from blood to bile is maintained by pumps of ABC superfamily secreting bile salts (bile acid dependent flow) and other solutes: bilirubin, phospholipids, glutathione and inorganic salts (bile acid independent flow).