In vitro evaluation of the transportability of viable primary human liver cells originating from discarded donor organs in bioreactors.
BACKGROUND
The use of primary human liver cells obtained from discarded donor organs is increasingly favored for cell-based extracorporeal liver support systems. However, as cryopreservation of primary human hepatocytes causes a significant loss of metabolic activity, the transport of bioreactors with viable liver cells is required. The aim of this study was to evaluate the impact of two major potential threats to viable cells during transport: temperature changes and mechanical stress.


METHODS
In each experiment three hollow fiber-based bioreactors were charged with primary human liver cells originating from the same discarded donor organ and were simultaneously kept under culture conditions for 8 days. In total, 18 bioreactors were evaluated. On the fifth day the bioreactors were exposed to hypothermia (4 degrees C, n = 3), to hyperthermia (42 degrees C, n = 3), or served as normothermic controls (37 degrees C, n = 3). In a second test series bioreactors were exposed to vibration (21 Hz for 20 min, thereafter 7 Hz for 160 min, n = 3), or were operated as control cultures (n = 6). The release of hepatocyte-specific enzymes was determined as an indicator for cell damage.


RESULTS
Hypothermic stress resulted in a significant release of transaminases and led to disturbances of the histological integrity, all indicating a high degree of cell damage. When compared with the control cultures, hyperthermia and mechanical stress in terms of vibration had no significant effect on the cells.


CONCLUSIONS
The transport of hollow fiber bioreactors charged with viable primary human liver cells appears to be feasible in transport monitors for perfusion and temperature control.
Sauer
Igor M
IM
General, Visceral and Transplantation Surgery, Charité, Campus Virchow Klinikum, Berlin, Germany. igor.sauer@charite.de
Schwartlander
Ruth
R
Van der Jagt
Olav
O
Steffen
Ingo
I
Efimova
Ekaterina
E
Pless
Gesine
G
Kehr
Daniel C
DC
Kardassis
Dimitrios
D
Fruhauf
Jan H
JH
Gerlach
Joerg C
JC
Neuhaus
Peter
P
D019149:Bioreactors; D002470:Cell Survival; D003080:Cold Temperature / Q000009:adverse effects; D005240:Feasibility Studies; D022781:Hepatocytes / Q000502:physiology*; D006358:Hot Temperature / Q000009:adverse effects; D006801:Humans; D009926:Organ Preservation / Q000379:methods*; D013048:Specimen Handling / Q000379:methods*; D013314:Stress, Mechanical; D009927:Tissue and Organ Procurement