AIM: To develop a hepatocyte cell line, we immortalized primary porcine hepatocytes with a retroviral vector SSR#69 containing the Simian Virus 40 T antigen (SV40T ag). METHODS: We first established a method of porcin...AIM: To develop a hepatocyte cell line, we immortalized primary porcine hepatocytes with a retroviral vector SSR#69 containing the Simian Virus 40 T antigen (SV40T ag). METHODS: We first established a method of porcine hepatocyte isolation with a modified four-step retrograde perfusion technique. Then the porcine hepatocytes were immortalized with retroviral vector SSR#69 expressing SV40T and hygromycin-resistance genes flanked by paired loxP recombination targets. SV40T cDNA in the expanded cells was subsequently excised by Cre/LoxP site-specific recombination. RESULTS: The resultant hepatocytes with high viability (97%) were successfully immortalized with retroviral vector SSR#69. One of the immortalized clones showed the typical morphological appearance, TJPH-1, and was selected by clone rings and expanded in culture. After excision of the SV40T gene with Cre-recombinase, cells stopped growing. The population of reverted cells exhibited the characteristics of differentiated hepatocytes. CONCLUSION: In conclusion, we herein describe a modified method of hepatocyte isolation and subsequently established a porcine hepatocyte cell line mediated by retroviral transfer and site-specific recombination.展开更多
Behavior of Ca/P ceramics enhanced by MgAl//2O//4 in simulated body fluid (SBF) was studied. Results show that, biological apatite can precipitate on the surface of Ca/P composite ceramics, however, dissolution of th...Behavior of Ca/P ceramics enhanced by MgAl//2O//4 in simulated body fluid (SBF) was studied. Results show that, biological apatite can precipitate on the surface of Ca/P composite ceramics, however, dissolution of the surface of Ca/P composite ceramics also occurs due to a kind of amorphous material which is produced through reaction of MgAl//2O//4 with Ca/P ceramics at elevated temperature; therefore, the behavior of Ca/P composite ceramics in SBF mainly depends on those two processes. Analysis of joint influence of those two processes show that low content of MgAl//2O//4 favors the formation of biological apatite, therefore, brings about higher bioactivity. (Edited author abstract) 23 Refs.展开更多
基金Supported by The Major Scientific and Technological Project of Hubei Province, No. 2007ABD005
文摘AIM: To develop a hepatocyte cell line, we immortalized primary porcine hepatocytes with a retroviral vector SSR#69 containing the Simian Virus 40 T antigen (SV40T ag). METHODS: We first established a method of porcine hepatocyte isolation with a modified four-step retrograde perfusion technique. Then the porcine hepatocytes were immortalized with retroviral vector SSR#69 expressing SV40T and hygromycin-resistance genes flanked by paired loxP recombination targets. SV40T cDNA in the expanded cells was subsequently excised by Cre/LoxP site-specific recombination. RESULTS: The resultant hepatocytes with high viability (97%) were successfully immortalized with retroviral vector SSR#69. One of the immortalized clones showed the typical morphological appearance, TJPH-1, and was selected by clone rings and expanded in culture. After excision of the SV40T gene with Cre-recombinase, cells stopped growing. The population of reverted cells exhibited the characteristics of differentiated hepatocytes. CONCLUSION: In conclusion, we herein describe a modified method of hepatocyte isolation and subsequently established a porcine hepatocyte cell line mediated by retroviral transfer and site-specific recombination.
文摘Behavior of Ca/P ceramics enhanced by MgAl//2O//4 in simulated body fluid (SBF) was studied. Results show that, biological apatite can precipitate on the surface of Ca/P composite ceramics, however, dissolution of the surface of Ca/P composite ceramics also occurs due to a kind of amorphous material which is produced through reaction of MgAl//2O//4 with Ca/P ceramics at elevated temperature; therefore, the behavior of Ca/P composite ceramics in SBF mainly depends on those two processes. Analysis of joint influence of those two processes show that low content of MgAl//2O//4 favors the formation of biological apatite, therefore, brings about higher bioactivity. (Edited author abstract) 23 Refs.
