17 research outputs found
Dried human skin fibroblasts as a new substratum for functional culture of hepatic cells
The primary hepatocytes culture is still one of the main challenges in toxicology studies in the drug discovery process, development of in vitro models to study liver function, and cell-based therapies. Isolated hepatocytes display a rapid decline in viability and liver-specific functions including albumin production, conversion of ammonia to urea, and activity of the drug metabolizing enzymes. A number of methods have been developed in order to maintain hepatocytes in their highly differentiated state in vitro. Optimization of culture conditions includes a variety of media formulations and supplements, growth surface coating with the components of extracellular matrix or with synthetic polymers, three-dimensional growth scaffolds and decellularized tissues, and coculture with other cell types required for the normal cell-cell interactions. Here we propose a new substratum for hepatic cells made by drying confluent human skin fibroblasts' culture. This growth surface coating, prepared using maximally simplified procedure, combines the advantages of the use of extracellular matrices and growth factors/cytokines secreted by the feeder layer cells. In comparison to the hepatoma cells grown on a regular tissue culture plastic, cells cultured on the dried fibroblasts were able to synthesize albumin in larger quantities and to form greater number of apical vacuoles. Unlike the coculture with the living feeder layer cells, the number of cells grown on the new substratum was not reduced after fourteen days of culture. This fact could make the dried fibroblasts coating an ideal candidate for the substrate for non-dividing human hepatocytes
Hollow fiber bioreactor with genetically modified hepatic cells as a model of biologically active function block of the bioartificial liver
Chronic liver disease and cirrhosis, that can lead to liver failure, are major public health issues, with liver
transplantation as the only effective treatment. However, the limited availability of transplantable organs has
spurred research into alternative therapies, including bioartificial livers. To date, liver hybrid support devices,
using porcine hepatocytes or hepatoma-derived cell lines, have failed to demonstrate efficacy in clinical trials.
Here, for the first time, we report the construction of a model of biologically active function block of bioartificial
liver based on a hollow fiber bioreactor populated with genetically modified hepatic cells. For
comprehensive comparison the culturing of hepatic cells was carried out in both static and dynamic conditions in
a medium that flowed through porous polysulfone capillaries. The most crucial parameters, such as cell viability,
glucose consumption, albumin secretion and urea production, were analyzed in static conditions while glucose
usage and albumin production were compared in dynamic cell cultures. This model has the potential to improve
the development of bioartificial liver devices and contribute to the treatment of patients with impaired liver
function
Populations of cells in liver isolates (light scatters).
<p>Exemplary flow cytometric dot plots showing populations of cells isolated from human liver tissue in light scatters: forward—FSC and side—SSC. Depending on specimen, there are minimum 2 and maximum 4 distinct cells populations (labeled P2 –P5) identified in the P1 gate (cellular debris and doublets excluded). Population P2 –blue dots; P3 –yellow dots; P4 –green dots; P5 –purple dots. The percentage of cells in the individual populations of the P1 gate—specimen H27-14: P2 = 35.9%, P3 = 62.8%; H26-14: P2 = 32.3%, P3 = 52.8%, P4 = 6.1%; H16-13: P2 = 47.6%, P3 = 46.4%, P4 = 1.7%, P5 = 1.3%; H22-13: P2 = 40.6%, P3 = 41.6%, P4 = 11%, P5 = 1.1%.</p
The characteristic of liver tissue used and the main results of liver cells isolations.
<p>The characteristic of liver tissue used and the main results of liver cells isolations.</p