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.
Department of Surgery, Klinikum Grosshadern, L.M. University of Munich, Germany.
H G
HG
Koebe
M
M
Wick
U
U
Cramer
V
V
Lange
F W
FW
Schildberg
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