Impact of Percoll purification on isolation of primary human hepatocytes
| Autor: | R. Horner, J. G. M. V. Gassner, M. Kluge, P. Tang, S. Lippert, K. H. Hillebrandt, S. Moosburner, A. Reutzel-Selke, J. Pratschke, I. M. Sauer, N. Raschzok |
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| Rok vydání: | 2019 |
| Předmět: |
centrifugation
Cell Survival lcsh:R human hepatocytes lcsh:Medicine Povidone Percoll purification Cell Separation Translational research Silicon Dioxide Article Preclinical research Hepatocytes Hepatectomy Humans lcsh:Q Aspartate Aminotransferases Biomaterials - cells lcsh:Science Cells Cultured 600 Technik Medizin angewandte Wissenschaften::610 Medizin und Gesundheit::610 Medizin und Gesundheit |
| Zdroj: | Scientific Reports Scientific Reports, Vol 9, Iss 1, Pp 1-10 (2019) |
| DOI: | 10.17169/refubium-25586 |
| Popis: | Research and therapeutic applications create a high demand for primary human hepatocytes. The limiting factor for their utilization is the availability of metabolically active hepatocytes in large quantities. Centrifugation through Percoll, which is commonly performed during hepatocyte isolation, has so far not been systematically evaluated in the scientific literature. 27 hepatocyte isolations were performed using a two-step perfusion technique on tissue obtained from partial liver resections. Cells were seeded with or without having undergone the centrifugation step through 25% Percoll. Cell yield, function, purity, viability and rate of bacterial contamination were assessed over a period of 6 days. Viable yield without Percoll purification was 42.4 × 106 (SEM ± 4.6 × 106) cells/g tissue. An average of 59% of cells were recovered after Percoll treatment. There were neither significant differences in the functional performance of cells, nor regarding presence of non-parenchymal liver cells. In five cases with initial viability of |
| Databáze: | OpenAIRE |
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