The Feasibility Study Of A Biomimetic 3-Dimentional Matrix For High-density Culture Of Swine Hepatocytes

Objective To study the feasibility of a biomimetic 3-Dimentinal culture matrix for high- density culture of swine hepatocytes by observing the function and morphological characteristic of the cultured cell.Methods Swine hepatocytes were isolated by two-step perfusion and cultured in a unique cell culture setting based on biomimetic 3-Dimentinal culture matrix four times. Observing the changes of AST,ALT,LDH, TBIL, albumin ,diazepam , urea concentration in setting and oxygen pressure difference between the exit and entry of setting at 0,2,4, 6, 8 hour after the beginning of culture, analyzing the diazepam clearance and urea(BUN) synthesizing rate during 0-2,2-4, 4-6,6-8 hour,observing morphological characteristics of hepatocytes in culture matrix. Results During the total period of 8 hours, there were no significant difference in concentration average of AST,ALT, TBIL,LDH and oxygen pressure difference average among different groups according to different time (P>0.05) ,but significant difference observed in concentration average of diazepam and urea (P<0.05) ; there were significant difference in diazepam clearance and urea(BUN) synthesizing rate among different groups according to different period (P<0.05) . The typical cellular morphological characteristics of hepatocytes could be observed in culture matrix. The density of cultured hepatocytes reached 10~7/ml.Conclusions The hepatocytes in culture matrix keep steady biochemical activity, good substance metabolic transformation and biosynthesis ability , typical cell physiological and morphological characteristics, this culture matrix could provide an environment that was more closely resembling that in vivo and achieved high density and activity culture of hepatocytes, which maybe more eligible for use in the study of bioartificial liver.