Function Test And Application Of Human Fetal Hepatocyte Line L-02

【OBJECTIVE】TO CONFIRM THE HEPATOCYTIC FUNCTION OF L-02 AND THE POTENTIAL TO TREAT ACUTE LIVER FAILURE.【METHODS】THE EXPRESSION OF ALBUMIN AND SOME HEPATOCYTE SPECIFIC MOLECULES IN L-02 CELLS WERE DETECTED BY WESTERN-BLOTTING, RT-PCR AND IMMUNOFLUORESCENCE. WE USED A RAT ALF MODEL INDUCED BY 90% HEPATECTOMY TO DETERMINE THE CELL FUNCTION. THE RATS IN GROUP 1 (G1) UNDERWENT L-02 CELLS TRANSPLANTATION INTO THE SPLEEN 1 DAY BEFORE HEPATECTOMY, AND GROUP 2 (G2) WITHOUT TRANSPLANTATION AS CONTROL. SURVIVAL RATE WAS RECORDED FOR EACH GROUP. THE SERUM SAMPLES WERE COLLECTED PER 24 HOURS FOR 7DAYS TO MEASURE BIOCHEMICAL PARAMETERS THAT INCLUDE TOTAL BILIRUBIN (T-BIL), GLUTAMIC-PYRUVIC TRANSAMINASE (ALT) AND BLOOD AMMONIA AND SO ON. TO EVALUATE TUMORIGENICITY OF L-02 CELLS, WE SUBCUTANEOSLY TRANSPLANTED THe cells into SCID mice, and tumor formation was observed for 4 weeks. Hepg-2 cells were used as a positive control.【Results】The expressions of albumin (ALB), HBCF-X (human blood coagulation factor X) and GST- (glutathione-S-transferase ) were detected in L-02 cells. And the amount of albumin expressed in L-02 cells was similar to that in human primary hepatocytes. Survival rate of G1 rats at 30 days was over 70%, whereas all rats in G2 died of acute liver failure within 72 hours. Biochemical parameters showed a significant improve in G1. The values reached the highest levels at 2 days postoperatively, and then presented a downtrend. Also there was no tumor formation when transplanted L-02 cells.【Conclusion】L-02 cells express the main molecules of human hepatocytes. More interestingly, they are able to effectively improve liver function and traumatically prolong the survival of ALF rats. These studies showed the L-02 cell line is a feasible cell source for hepatocytes transplantation and BAL.