Abstract: Objective To explore whether the function of the CL-1 hepatocytes, co-cultured with human hepatic stellate cells
(HSC) on microearriers was better than that cultured without HSC. Methods CL-1 hepatocytes were divided into 2 groups.
The co-culture group was cultured with HSC in DMEM culture medium containing 10% fetal bovine serum, and HSC were not
added in single-culture group. The cytomorphology was observed by inverted microscope and scanning electron microscopy.
The effects of different culture method on the proliferation in vitro were analyzed by MTT assay. The function of hepatocytes
was evaluated through measuring the concentration of ALT and albumin. Results The inverted microscope and the MTT
staining results showed that the quantity and viability of the human hepatocyte (C3A) in bidirectional bioreactor group were
much better than the RCMW group. The growth curve results showed that the density of the human hepatocyte (C3A) was
firstly increased and then decreased in both groups, and the peak of the curve appeared in day 5. The density of human liver
cells in the second generation of bi-directional rotation and perfusion microgravity bioreactor group was significantly higher
than the RCMW group from day1 to day 7 (P<0.05). The functional results showed that the albumin and urea concentration,
which reached the peak on day 5, also gone up firstly and then gradually gone down in both teams. And the albumin and urea
concentration in bidirectional bioreactor group was significantly higher than RCMW group from day 1 to day7 (P<0.01).
Besides, the concentration of ALT and AST in bidirectional bioreactor group was significantly lower than RCMW group from
day 1 to day 7 (P<0.05). Conclusions This study shows that this new culture method is advantageous in enhancing
cell viability and function. It indicates that co-culturing hepatocytes with HSC has a good application prospect in the
development of artificial liver technology.