Construction of IL-13 Gene-Modified Rattus Hepatic Stem Cells_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

CHEN Gang ¹ , HAN Jiang ² , LIU Yongkang ¹ , AI Junhua ¹ , DONG Jiahong ¹

1.Institute of Hepatobiliary Surgery, Southwest Hospital， The Third Military Medical University, Chongqing 400038, China;;
2.Department of Hepatobiliary Surgery, Kunming General Hospital of PLA, Chengdu Military Region, Kunming 650032, ChinaCorresponding Author: DONG Jia-hong, E-mail: dongjh301@163.com;

Corresponding author：

Keywords：

Lentivirus; Interleukin-13; Hepatic oval cell; Hepatic stem cell; Vector construction

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

Objective 　To construct gene-modified hepatic stem cells (WB-F344 cells), which have rat IL-13 gene and can secrete the recombinant rat IL-13 cytokine in the cells.
Methods 　Firstly, the rat IL-13 sequences were synthesized. Then the sequences were amplificated in bacterium coli after recombinated with pWPXL-MOD plasmid. After PCR and sequence identification, the positive clones were packaged into lentivirus. After detecting the virus titer, the WB-F344 cells with constructed lentivirus vector with rat IL-13 gene were cultured, then the valid targets (expression level of the IL-13) were detected by real time-PCR and Western blot in cultured WB-F344 cells on 5 days.
Results 　The valid DNA of rat IL-13 was recombinated and packaged in lentivirus vector. The recombinant gene sequence was correct by checking with gene sequence test. Then the recombinant was introducted into the WB-F344 cells cultures. The best multiplicity of infection (MOI) value for effective transfection was 5. IL-13 had been detected on day 5 after transfection by checking with real-time PCR and Western blot.
Conclusion 　The recombinant rat IL-13 gene with lentivirus vector is constructed and gene-modified WB-F344 cells are cultured successfully, which can be used in next animal experiment.

Citation： CHEN Gang,HAN Jiang,LIU Yongkang,AI Junhua,DONG Jiahong. Construction of IL-13 Gene-Modified Rattus Hepatic Stem Cells. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2009, 16(4): 276-280. doi: Copy

1.	Alison MR, Vig P, Russo F, et al. Hepatic stem cells: from inside and outside the liver? [Ｊ]. Cell Prolif, 2004; 37(1): 1-21.
2.	Kafri T, van Praag H, Gage FH, et al. Lentiviral vectors: regulated gene expression [Ｊ]. Mol Ther, 2000; 1(6): 516-521.
3.	Xu K, Ma H, McCown TJ, et al. Generation of a stable cell line producing high-titer self-inactivating lentiviral vectors [Ｊ]. Mol Ther, 2001; 3(1): 97-104.
4.	Charrier S, Stockholm D, Seye K, et al. A lentiviral vector encoding the human Wiskott-Aldrich syndrome protein corrects immune and cytoskeletal defects in WASP knockout mice [Ｊ]. Gene Ther, 2005; 12(7): 597-606.
5.	Trono D. Lentiviral vectors: turning a deadly foe into a therapeutic agent [Ｊ]. Gene Ther, 2000; 7(1): 20-23.
6.	Sinn PL, Sauter SL, McCray PB Jr. Gene therapy progress and prospects: development of improved lentiviral and retroviral vectors—design, biosafety, and production [Ｊ]. Gene Ther， 2005; 12(14): 1089-1098.
7.	Romano G. Current development of lentiviral-mediated gene transfer [Ｊ]. Drug News Perspect， 2005; 18(2): 128-134.
8.	Zufferey R, Nagy D, Mandel RJ, et al. Multiply attenuated lentiviral vector achieves efficient gene delivery in vivo [Ｊ]. Nat Biotechnol， 1997; 15(9): 871-875.
9.	Naldini L, Blmer U, Gage FH, et al. Efficient transfer, integration, and sustained long-term expression of the transgene in adult rat brains injected with a lentiviral vector [Ｊ]. Proc Natl Acad Sci USA， 1996; 93(21): 11382-11388.
10.	Miyoshi H, Smith KA, Mosier DE, et al. Transduction of human CD34+ cells that mediate long-term engraftment of NOD/SCID mice by HIV vectors [Ｊ]. Science， 1999; 283(5402): 682-686.
11.	Kafri T, Blmer U, Peterson DA, et al. Sustained expression of genes delivered directly into liver and muscle by lentiviral vectors [Ｊ]. Nat Genet， 1997; 17(3): 314-317.
12.	Chomarat P, Banchereau J. Interleukin-4 and interleukin-13: their similarities and discrepancies [Ｊ]. Int Rev Immunol, 1998; 17(1-4): 1-52.
13.	Gorczynski RM, Cohen Z, Fu XM, et al. Interleukin-13, in combination with anti-interleukin-12, increases graft prolongation after portal venous immunization with cultured allogeneic bone marrow-derived dendritic cells [Ｊ]. Transplantation, 1996; 62(11): 1592-1600.
14.	陈刚, 傅志仁, 王梁华, 等. 人白介素-13对牛主动脉内皮细胞表达黏附分子的影响 [Ｊ]. 中国普外基础与临床杂志, 2005；　12(5): 469-472.
15.	陈刚, 晋云, 李雪松, 等. 保护性基因A20在血管内皮细胞中的诱导表达 [Ｊ]. 细胞与分子免疫学杂志, 2007; 23(12): 1200-1202.
16.	Qin AL, Zhou XQ, Zhang W, et al. Characterization and enrichment of hepatic progenitor cells in adult rat liver [Ｊ]. World J Gastroenterol, 2004; 10(10): 1480-1486.
17.	Strain AJ, Crosby HA, Nijjar S, et al. Human liver-derived stem cells [Ｊ]. Semin Liver Dis, 2003; 23(4): 373-384.
18.	Wang X, Foster M, Al-Dhalimy M, et al. The origin and liver repopulating capacity of murine oval cells [Ｊ]. Proc Natl Acad Sci USA, 2003; 100 Suppl 1: 11881-11888.
19.	Sell S. Heterogeneity and plasticity of hepatocyte lineage cells [Ｊ]. Hepatology, 2001; 33(3): 738-750.
20.	Oh SH, Hatch HM, Petersen BE. Hepatic oval ‘stem’ cell in liver regeneration [Ｊ]. Semin Cell Dev Biol, 2002; 13(6): 405-409.
21.	Muller-Borer BJ, Cascio WE, Anderson PA, et al. Adult-derived liver stem cells acquire a cardiomyocyte structural and functional phenotype ex vivo [Ｊ]. Am J Pathol, 2004; 165(1): 135-145.
22.	Chramostová K, Vondrácek J, Sindlerová L, et al. Polycyclic aromatic hydrocarbons modulate cell proliferation in rat hepatic epithelial stem-like WB-F344 cells [Ｊ]. Toxicol Appl Pharmacol, 2004; 196(1): 136-148.
23.	Suzuki A, Nakauchi H, Taniguchi H. In vitro production of functionally mature hepatocytes from prospectively isolated hepatic stem cells [Ｊ]. Cell Transplant, 2003; 12(5): 469-473.
24.	Poulsom R, Alison MR, Forbes SJ, et al. Adult stem cell plasticity [Ｊ]. J Pathol, 2002; 197(4): 441-456.
25.	Yin L, Sun M, Ilic Z, et al. Derivation, characterization, and phenotypic variation of hepatic progenitor cell lines isolated from adult rats [Ｊ]. Hepatology, 2002; 35(2): 315-324.
26.	〖ＪＰ３〗Zheng YW, Taniguchi H. Diversity of hepatic stem cells in the fetal and adult liver [Ｊ]. Semin Liver Dis, 2003; 23(4): 337-348.
27.	Weber A. Immortalization of hepatic progenitor cells [Ｊ]. Pathol Biol (Paris), 2004; 52(2): 93-96.

1. Alison MR, Vig P, Russo F, et al. Hepatic stem cells: from inside and outside the liver? [Ｊ]. Cell Prolif, 2004; 37(1): 1-21.
2. Kafri T, van Praag H, Gage FH, et al. Lentiviral vectors: regulated gene expression [Ｊ]. Mol Ther, 2000; 1(6): 516-521.
3. Xu K, Ma H, McCown TJ, et al. Generation of a stable cell line producing high-titer self-inactivating lentiviral vectors [Ｊ]. Mol Ther, 2001; 3(1): 97-104.
4. Charrier S, Stockholm D, Seye K, et al. A lentiviral vector encoding the human Wiskott-Aldrich syndrome protein corrects immune and cytoskeletal defects in WASP knockout mice [Ｊ]. Gene Ther, 2005; 12(7): 597-606.
5. Trono D. Lentiviral vectors: turning a deadly foe into a therapeutic agent [Ｊ]. Gene Ther, 2000; 7(1): 20-23.
6. Sinn PL, Sauter SL, McCray PB Jr. Gene therapy progress and prospects: development of improved lentiviral and retroviral vectors—design, biosafety, and production [Ｊ]. Gene Ther， 2005; 12(14): 1089-1098.
7. Romano G. Current development of lentiviral-mediated gene transfer [Ｊ]. Drug News Perspect， 2005; 18(2): 128-134.
8. Zufferey R, Nagy D, Mandel RJ, et al. Multiply attenuated lentiviral vector achieves efficient gene delivery in vivo [Ｊ]. Nat Biotechnol， 1997; 15(9): 871-875.
9. Naldini L, Blmer U, Gage FH, et al. Efficient transfer, integration, and sustained long-term expression of the transgene in adult rat brains injected with a lentiviral vector [Ｊ]. Proc Natl Acad Sci USA， 1996; 93(21): 11382-11388.
10. Miyoshi H, Smith KA, Mosier DE, et al. Transduction of human CD34+ cells that mediate long-term engraftment of NOD/SCID mice by HIV vectors [Ｊ]. Science， 1999; 283(5402): 682-686.
11. Kafri T, Blmer U, Peterson DA, et al. Sustained expression of genes delivered directly into liver and muscle by lentiviral vectors [Ｊ]. Nat Genet， 1997; 17(3): 314-317.
12. Chomarat P, Banchereau J. Interleukin-4 and interleukin-13: their similarities and discrepancies [Ｊ]. Int Rev Immunol, 1998; 17(1-4): 1-52.
13. Gorczynski RM, Cohen Z, Fu XM, et al. Interleukin-13, in combination with anti-interleukin-12, increases graft prolongation after portal venous immunization with cultured allogeneic bone marrow-derived dendritic cells [Ｊ]. Transplantation, 1996; 62(11): 1592-1600.
14. 陈刚, 傅志仁, 王梁华, 等. 人白介素-13对牛主动脉内皮细胞表达黏附分子的影响 [Ｊ]. 中国普外基础与临床杂志, 2005；　12(5): 469-472.
15. 陈刚, 晋云, 李雪松, 等. 保护性基因A20在血管内皮细胞中的诱导表达 [Ｊ]. 细胞与分子免疫学杂志, 2007; 23(12): 1200-1202.
16. Qin AL, Zhou XQ, Zhang W, et al. Characterization and enrichment of hepatic progenitor cells in adult rat liver [Ｊ]. World J Gastroenterol, 2004; 10(10): 1480-1486.
17. Strain AJ, Crosby HA, Nijjar S, et al. Human liver-derived stem cells [Ｊ]. Semin Liver Dis, 2003; 23(4): 373-384.
18. Wang X, Foster M, Al-Dhalimy M, et al. The origin and liver repopulating capacity of murine oval cells [Ｊ]. Proc Natl Acad Sci USA, 2003; 100 Suppl 1: 11881-11888.
19. Sell S. Heterogeneity and plasticity of hepatocyte lineage cells [Ｊ]. Hepatology, 2001; 33(3): 738-750.
20. Oh SH, Hatch HM, Petersen BE. Hepatic oval ‘stem’ cell in liver regeneration [Ｊ]. Semin Cell Dev Biol, 2002; 13(6): 405-409.
21. Muller-Borer BJ, Cascio WE, Anderson PA, et al. Adult-derived liver stem cells acquire a cardiomyocyte structural and functional phenotype ex vivo [Ｊ]. Am J Pathol, 2004; 165(1): 135-145.
22. Chramostová K, Vondrácek J, Sindlerová L, et al. Polycyclic aromatic hydrocarbons modulate cell proliferation in rat hepatic epithelial stem-like WB-F344 cells [Ｊ]. Toxicol Appl Pharmacol, 2004; 196(1): 136-148.
23. Suzuki A, Nakauchi H, Taniguchi H. In vitro production of functionally mature hepatocytes from prospectively isolated hepatic stem cells [Ｊ]. Cell Transplant, 2003; 12(5): 469-473.
24. Poulsom R, Alison MR, Forbes SJ, et al. Adult stem cell plasticity [Ｊ]. J Pathol, 2002; 197(4): 441-456.
25. Yin L, Sun M, Ilic Z, et al. Derivation, characterization, and phenotypic variation of hepatic progenitor cell lines isolated from adult rats [Ｊ]. Hepatology, 2002; 35(2): 315-324.
26. 〖ＪＰ３〗Zheng YW, Taniguchi H. Diversity of hepatic stem cells in the fetal and adult liver [Ｊ]. Semin Liver Dis, 2003; 23(4): 337-348.
27. Weber A. Immortalization of hepatic progenitor cells [Ｊ]. Pathol Biol (Paris), 2004; 52(2): 93-96.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Construction of IL-13 Gene-Modified Rattus Hepatic Stem Cells

Abstract Full text Figures/Tables Video References Cited by

Format

Content