Summary
Transdifferentiation is defined as an irreversible switch in postnatal life of one differentiated cell to another. Transdifferentiation from different cellular origins into pancreatic-like β-cells is of clinical significance since this approach may offer a potential cure for diabetes. In order to achieve this goal, the liver is considered as a suitable candidate due to its close developmental relationship to the pancreas, its large size and a well-documented regenerative capacity that could provide enough original tissues to initiate the transdifferentiation procedure. In this chapter, we describe a protocol to overexpress Pdx1, a master regulator essential for pancreas development in the cultured human liver cell line, HepG2.
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References
Slack, J.M., (1985). Homoeotic transformations in man: implications for the mechanism of embryonic development and for the organization of epithelia. J Theor Biol, 114: 463–490
Slack, J.M., (1986). Epithelial metaplasia and the second anatomy. Lancet, 2: 268–271
Tosh, D. and J.M. Slack, (2002). How cells change their phenotype. Nat Rev Mol Cell Biol, 3: 187–194
Burke, Z.D. and D. Tosh, (2005). Therapeutic potential of transdifferentiated cells. Clin Sci (Lond), 108: 309–321
Kume, S., (2005). Stem-cell-based approaches for regenerative medicine. Dev Growth Differ, 47: 393–402
Slack, J.M. and D. Tosh, (2001). Transdifferentiation and metaplasia - switching cell types. Curr Opin Genet Dev, 11: 581–586
Lardon, J., N. Huyens, I. Rooman, and L. Bouwens, (2004). Exocrine cell transdifferentiation in dexamethasone-treated rat pancreas. Virchows Arch, 444: 61–65
Baeyens, L., S. De Breuck, J. Lardon, J.K. Mfopou, I. Rooman, and L. Bouwens, (2005). In vitro generation of insulin-producing beta cells from adult exocrine pancreatic cells. Diabetologia, 48: 49–57
Bonner-Weir, S., M. Taneja, G.C. Weir, K. Tatarkiewicz, K.H. Song, A. Sharma, and J.J. O’Neil, (2000). In vitro cultivation of human islets from expanded ductal tissue. Proc Natl Acad Sci U S A, 97: 7999–8004
Yatoh, S., R. Dodge, T. Akashi, A. Omer, A. Sharma, G.C. Weir, and S. Bonner-Weir, (2007). Differentiation of affinity-purified human pancreatic duct cells to beta-cells. Diabetes, 56: 1802–1809
Yamada, S., Y. Yamamoto, M. Nagasawa, A. Hara, T. Kodera, and I. Kojima, (2006). In vitro transdifferentiation of mature hepatocytes into insulin-producing cells. Endocr J, 53: 789–795
Ferber, S., A. Halkin, H. Cohen, I. Ber, Y. Einav, I. Goldberg, I. Barshack, R. Seijffers, J. Kopolovic, N. Kaiser, and A. Karasik, (2000). Pancreatic and duodenal homeobox gene 1 induces expression of insulin genes in liver and ameliorates streptozotocin-induced hyperglycemia. Nat Med, 6: 568–572
Wells, J.M. and D.A. Melton, (1999). Vertebrate endoderm development. Annu Rev Cell Dev Biol, 15: 393–410
Jung, J., M. Zheng, M. Goldfarb, and K.S. Zaret, (1999). Initiation of mammalian liver development from endoderm by fibroblast growth factors. Science, 284: 1998–2003
Murtaugh, L.C., (2007). Pancreas and beta-cell development: from the actual to the possible. Development, 134: 427–438
Offield, M.F., T.L. Jetton, P.A. Labosky, M. Ray, R.W. Stein, M.A. Magnuson, B.L. Hogan, and C.V. Wright, (1996). PDX-1 is required for pancreatic outgrowth and differentiation of the rostral duodenum. Development, 122: 983–995
Holland, A.M., L.J. Gonez, G. Naselli, R.J. Macdonald, and L.C. Harrison, (2005). Conditional expression demonstrates the role of the homeodomain transcription factor Pdx1 in maintenance and regeneration of beta-cells in the adult pancreas. Diabetes, 54: 2586–2595
Ahlgren, U., J. Jonsson, L. Jonsson, K. Simu, and H. Edlund, (1998). beta-cell-specific inactivation of the mouse Ipf1/Pdx1 gene results in loss of the beta-cell phenotype and maturity onset diabetes. Genes Dev, 12: 1763–1768
Jonsson, J., L. Carlsson, T. Edlund, and H. Edlund, (1994). Insulin-promoter-factor 1 is required for pancreas development in mice. Nature, 371: 606–609
Ber, I., K. Shternhall, S. Perl, Z. Ohanuna, I. Goldberg, I. Barshack, L. Benvenisti-Zarum, I. Meivar-Levy, and S. Ferber, (2003). Functional, persistent, and extended liver to pancreas transdifferentiation. J Biol Chem, 278: 31950–31957
Miyatsuka, T., H. Kaneto, Y. Kajimoto, S. Hirota, Y. Arakawa, Y. Fujitani, Y. Umayahara, H. Watada, Y. Yamasaki, M.A. Magnuson, J. Miyazaki, and M. Hori, (2003). Ectopically expressed PDX-1 in liver initiates endocrine and exocrine pancreas differentiation but causes dysmorphogenesis. Biochem Biophys Res Commun, 310: 1017–1025
Tang, D.Q., L.Z. Cao, W. Chou, L. Shun, C. Farag, M.A. Atkinson, S.W. Li, L.J. Chang, and L.J. Yang, (2006). Role of Pax4 in Pdx1-VP16-mediated liver-to-endocrine pancreas transdifferentiation. Lab Invest, 86: 829–841
Li, W.C., M.E. Horb, D. Tosh, and J.M. Slack, (2005). In vitro transdifferentiation of hepatoma cells into functional pancreatic cells. Mech Dev, 122: 835–847
Aden, D.P., A. Fogel, S. Plotkin, I. Damjanov, and B.B. Knowles, (1979). Controlled synthesis of HBsAg in a differentiated human liver carcinoma-derived cell line. Nature, 282: 615–616
Dashti, N. and G. Wolfbauer, (1987). Secretion of lipids, apolipoproteins, and lipoproteins by human hepatoma cell line, HepG2: effects of oleic acid and insulin. J Lipid Res, 28: 423–436
Horb, M.E., C.N. Shen, D. Tosh, and J.M. Slack, (2003). Experimental Conversion of liver to pancreas. Curr Biol, 13: 105–115
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Li, WC. (2009). In Vitro Transdifferentiation of Human Hepatoma Cells into Pancreatic-Like Cells. In: Stocker, C. (eds) Type 2 Diabetes. Methods in Molecular Biology, vol 560. Humana Press. https://doi.org/10.1007/978-1-59745-448-3_9
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DOI: https://doi.org/10.1007/978-1-59745-448-3_9
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