Abstract
The aim of this study was to establish a stand-alone, perfused, rotary cell culture system using small human hepatocytes (SH) for bioartificial liver (BAL) support. SH were grown on cytodex 3 microcarriers (beads) to a maximum density of 1.2 ± 0.3 × 107 cells per mL within 12 days. Size of aggregates formed by up to 15 beads was regulated by rotation speed. Cell function was proven by treatment with ammonia and galactose, and metabolism was analyzed. Treatment strategy was comprised of two phases, namely growth phase and treatment phase. Cells were grown for 6 days and subsequently incubated with ammonia or galactose for 2 days, followed by a 2-day regeneration period and another 2-day treatment phase. Consumption of glucose, release of lactate dehydrogenase, formation of lactate, and production of urea and albumin were determined regularly. Mean galactose consumption was 50 μg per 106 cells per hour, ammonia-induced urea formation was 3.6 μg per 106 cells per hour, and albumin production was 110ng per 106 cells per hour. All metabolic parameters followed a logarithmic trend and were found to be very stable in the second half of the culture period when cells were treated with ammonia or galactose. Dissolved oxygen (%DO), pH, and temperature were monitored in-stream at intervals of 7 min, and the values were logged. Viability and morphology of cells were monitored via confocal microscopy. Viability was around 95% in controls and 90% during treatment. Promising results were obtained in support of our ongoing efforts to establish a fully autonomous BAL support device utilizing SH as a bridge to transplantation.
| Original language | English |
|---|---|
| Pages (from-to) | 1063-1073 |
| Number of pages | 11 |
| Journal | Tissue Engineering - Part A |
| Volume | 15 |
| Issue number | 5 |
| DOIs | |
| State | Published - May 1 2009 |
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ASJC Scopus subject areas
- Bioengineering
- Biochemistry
- Biomedical Engineering
- Biomaterials
Cite this
Introduction of a novel prototype bioartificial liver support system utilizing small human hepatocytes in rotary culture. / Wurm, Martin; Lubei, Verena; Caronna, Marco; Hermann, Martin; Buttiglieri, Stefano; Bodamer, Olaf; Muehl, Adolf; Tetta, Ciro; Margreiter, Raimund; Hengster, Paul.
In: Tissue Engineering - Part A, Vol. 15, No. 5, 01.05.2009, p. 1063-1073.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Introduction of a novel prototype bioartificial liver support system utilizing small human hepatocytes in rotary culture
AU - Wurm, Martin
AU - Lubei, Verena
AU - Caronna, Marco
AU - Hermann, Martin
AU - Buttiglieri, Stefano
AU - Bodamer, Olaf
AU - Muehl, Adolf
AU - Tetta, Ciro
AU - Margreiter, Raimund
AU - Hengster, Paul
PY - 2009/5/1
Y1 - 2009/5/1
N2 - The aim of this study was to establish a stand-alone, perfused, rotary cell culture system using small human hepatocytes (SH) for bioartificial liver (BAL) support. SH were grown on cytodex 3 microcarriers (beads) to a maximum density of 1.2 ± 0.3 × 107 cells per mL within 12 days. Size of aggregates formed by up to 15 beads was regulated by rotation speed. Cell function was proven by treatment with ammonia and galactose, and metabolism was analyzed. Treatment strategy was comprised of two phases, namely growth phase and treatment phase. Cells were grown for 6 days and subsequently incubated with ammonia or galactose for 2 days, followed by a 2-day regeneration period and another 2-day treatment phase. Consumption of glucose, release of lactate dehydrogenase, formation of lactate, and production of urea and albumin were determined regularly. Mean galactose consumption was 50 μg per 106 cells per hour, ammonia-induced urea formation was 3.6 μg per 106 cells per hour, and albumin production was 110ng per 106 cells per hour. All metabolic parameters followed a logarithmic trend and were found to be very stable in the second half of the culture period when cells were treated with ammonia or galactose. Dissolved oxygen (%DO), pH, and temperature were monitored in-stream at intervals of 7 min, and the values were logged. Viability and morphology of cells were monitored via confocal microscopy. Viability was around 95% in controls and 90% during treatment. Promising results were obtained in support of our ongoing efforts to establish a fully autonomous BAL support device utilizing SH as a bridge to transplantation.
AB - The aim of this study was to establish a stand-alone, perfused, rotary cell culture system using small human hepatocytes (SH) for bioartificial liver (BAL) support. SH were grown on cytodex 3 microcarriers (beads) to a maximum density of 1.2 ± 0.3 × 107 cells per mL within 12 days. Size of aggregates formed by up to 15 beads was regulated by rotation speed. Cell function was proven by treatment with ammonia and galactose, and metabolism was analyzed. Treatment strategy was comprised of two phases, namely growth phase and treatment phase. Cells were grown for 6 days and subsequently incubated with ammonia or galactose for 2 days, followed by a 2-day regeneration period and another 2-day treatment phase. Consumption of glucose, release of lactate dehydrogenase, formation of lactate, and production of urea and albumin were determined regularly. Mean galactose consumption was 50 μg per 106 cells per hour, ammonia-induced urea formation was 3.6 μg per 106 cells per hour, and albumin production was 110ng per 106 cells per hour. All metabolic parameters followed a logarithmic trend and were found to be very stable in the second half of the culture period when cells were treated with ammonia or galactose. Dissolved oxygen (%DO), pH, and temperature were monitored in-stream at intervals of 7 min, and the values were logged. Viability and morphology of cells were monitored via confocal microscopy. Viability was around 95% in controls and 90% during treatment. Promising results were obtained in support of our ongoing efforts to establish a fully autonomous BAL support device utilizing SH as a bridge to transplantation.
UR - http://www.scopus.com/inward/record.url?scp=66249088717&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=66249088717&partnerID=8YFLogxK
U2 - 10.1089/ten.tea.2008.0217
DO - 10.1089/ten.tea.2008.0217
M3 - Article
C2 - 18847359
AN - SCOPUS:66249088717
VL - 15
SP - 1063
EP - 1073
JO - Tissue Engineering - Part A
JF - Tissue Engineering - Part A
SN - 1937-3341
IS - 5
ER -