Preparation of functionally active immobilized and perfused mammalian cells: an example of the hepatocyte bioreactor.
In the present study, a method has been employed for hepatocyte immobilization in agarose threads which allows for cell perfusion. The rat hepatocytes are isolated from the liver. A 1.8% low-gelling agarose solution is prepared in warm Krebs-Henseleit solution. The agarose solution is mixed 1:1 with the hepatocytes and the cells are immobilized in agarose threads by extruding the agarose-cell mixture through cooled Chemfluor teflon (TFE) tubing. Light and electron microscopy studies indicated the integrity of the hepatocytes in the gel matrix. This system allows for liver cell perfusion and viability studies to be carried out non-invasively on the cells and provides data that are comparable to those obtained with a perfused isolated liver. Immobilized hepatocytes are an in vitro system worthy of further evaluation which may be useful in the studies of liver cell metabolism and the response of the liver to foreign chemicals.
Farghali
H
H
Institute of Pharmacology, First Medical Faculty, Charles University, Prague, Czech Republic.
Kameníková
L
L
Martínek
J
J
Lincová
D
D
Hynie
S
S
D000818:Animals; D001709:Biotechnology / Q000295:instrumentation* / Q000379:methods*; D002469:Cell Separation / Q000295:instrumentation* / Q000379:methods*; D005947:Glucose; D008099:Liver / Q000166:cytology* / Q000648:ultrastructure; D008854:Microscopy, Electron; D010477:Perfusion / Q000295:instrumentation* / Q000379:methods*; D051381:Rats; D012685:Sepharose; D014325:Tromethamine
C074097:Krebs-Henseleit solution; D014325:Tromethamine; D012685:Sepharose; D005947:Glucose