Collagen gel immobilisation provides a suitable cell matrix for long term human hepatocyte cultures in hybrid reactors.
An easy to apply culture technique is presented that protects a monolayer configuration of liver cells within an extracellular matrix. The Immobilising Gel (IG)-Technique not only preserves hepatocyte morphology and supports a variety of differentiated cell functions over long term periods, but also offers higher resistance of IG-culture systems against shear forces of fluids in a hybrid reactor device, as compared to other culture techniques. Human hepatocyte cultures in IG-Technique: DNA-normalised levels for the total production of cholinesterase, albumin, urea and lactate remained high throughout the investigational period (50 days). Glutamic-Pyruvic-Transaminase (GPT) release decreased after peak values during early culture adaptation. Electron Microscopic (EM) findings after the shear forces experiment revealed undisturbed subcellular structures and a preserved intercellular morphology, including bile canaliculi and desmosomes. We conclude that the IG-technique is of considerable advantage as compared to other culture systems, especially in the field of dynamic applications, e.g. hybrid reactors for artificial organ development.
Koebe
H G
HG
Department of Surgery, Klinikum Grosshadern, L.M. University of Munich, Germany.
Wick
M
M
Cramer
U
U
Lange
V
V
Schildberg
F W
FW
D000410:Alanine Transaminase / Q000378:metabolism; D000418:Albumins / Q000378:metabolism; D001187:Artificial Organs; D002478:Cells, Cultured; D003094:Collagen; D003584:Cytological Techniques; D004247:DNA / Q000378:metabolism; D005109:Extracellular Matrix; D005782:Gels; D006801:Humans; D006822:Hybrid Cells; D008099:Liver / Q000166:cytology*; D013997:Time Factors; D014021:Tissue Preservation / Q000379:methods*
D000418:Albumins; D005782:Gels; D003094:Collagen; D004247:DNA; D000410:Alanine Transaminase