Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
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Effect of Inhibitor of Glycogen Synthase Kinase 3 on Self-Renewal of Human Embryonic Stem Cells

  • Lee Eunyoung (Laboratory of Development and Differentiation, Korea Research Institute of Bioscience and Biotechnology(KRIBB)) ;
  • Rho Jeung-yon (Laboratory of Development and Differentiation, Korea Research Institute of Bioscience and Biotechnology(KRIBB)) ;
  • Yu Kwon (Laboratory of Development and Differentiation, Korea Research Institute of Bioscience and Biotechnology(KRIBB)) ;
  • Paik Sang-Gi (Department of Biology, Chungnam National University) ;
  • Lee Kyung-Kwang (Laboratory of Development and Differentiation, Korea Research Institute of Bioscience and Biotechnology(KRIBB)) ;
  • Han Yong-Mahn (Laboratory of Development and Differentiation, Korea Research Institute of Bioscience and Biotechnology(KRIBB))
  • Published : 2005.06.01
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Abstract

Human embryonic stem cells (hESCs) derived from the inner cell mass of blastocysts have the ability to renew themselves and to differentiate into cell types of all lineage. The present study was carried out to investigate whether the Wnt signaling pathway is related to maintaining self-renewal of hESCs. Glycogen Synthase Kinase 3 (GSK-3) inhibitor, BIO ((2'Z,3'E)-6-Bromoindirubin-3'-oxime) was treated to Miz-hES1 line for activation of Wnt signaling pathway. BIO-nontreated hESCs (control) and BID-treated hESCs were cultured for 5 days in the modified feeder-free system. During the culture of hESCs, differences were observed in the colony morphology between 2 groups. Controls were spread outwards whereas BIO-nontreated hESCs were clumped in the center and the differentiated cells were spreading outwards in the edges. The results of stem cell specific marker staining indicated that control were differentiated in large part whereas BIO-treated hESCs maintain self-renewal in the center of the colony. The results of lineage marker staining suggested that outer cells of the hESC colony were differentiated to the neuronal progenitor cells in both control and BIO-treated hESC. These results indicate that Wnt signaling is related to self-renewal in hESCs. In addition, control group showed higher composition of apoptotic cells $(23.76\%)$ than the BID-treated group $(5.59\%)$. These results indicate that BIO is effective on antapoptosis of hESCs.

Keywords

References

  1. Avilion AA, Nicolis SK, Pevny LH, Perez L, Vivian N, Lovell R (2003): Multipotent cell lineages in early mouse development depend on SOX2 function. Genes & Dev 17:126-140 https://doi.org/10.1101/gad.224503
  2. Cadigan KM, Nusse R (1997): Wnt signaling: a common theme in animal development. Genes Dev. 11:3286-305 https://doi.org/10.1101/gad.11.24.3286
  3. Chambers I, Colby D, Robertson M, Nichols J, Lee S, Tweedie S, Smith A (2003): Functional expression cloning of nanog, a pluripotency sustaining factor in embryonic stem cells. Cell 113:643-655 https://doi.org/10.1016/S0092-8674(03)00392-1
  4. Chenn A, Walsh CA (2002): Regulation of cerebral cortical size by control of cell cycle exit in neural precursors. Science 297:365-369 https://doi.org/10.1126/science.1074192
  5. Gat U, DasGupta R, Degenstein L, Fuchs E (1998): De novo hair follicle morphogenesis and hair tumors in mice expressing a truncated $\beta$-catenin in skin. Cell 95:605-614 https://doi.org/10.1016/S0092-8674(00)81631-1
  6. Hori Y (2002): Growth inhibitors promote differentiation of insulin-producing tissue from embryonic stem cells. Proc Natl Acad Sci USA 99:16105-16110
  7. Jamora C, Fuchs E (2002): Intracellular adhesion signaling and the cytoskeleton. Nature Cell Biol 4:E101 https://doi.org/10.1038/ncb0402-e101
  8. Kielman MF, Rindapaa M, Gaspar C, van Poppel N, Breukel C, van Leeuwen, S, Taketo MM, Roberts S, Smits R, Fodde R (2002): Apc modulates embryonic stem-cell differentiation by controlling the dosage of $\beta$-catenin signaling. Nature Genet 32:594-605 https://doi.org/10.1038/ng1045
  9. Kim JH, Auerbach JM, Rodriguez-Gomez JA, Velasco I, Gavin D, Lumelsky N, Lee SH, Nguyen J, Sanchez-Pernaute R, Bankiewicz K, McKay R (2002): Dopamine neurons derived from embryonic stem cellsin an animal model of Parkinson's disease. Nature 418:50-56 https://doi.org/10.1038/nature00900
  10. King TD, Jope RS (2005): Inhibition of glycogen synthase kinase-3 protects cells from intrinsic but not extrinsic oxidative stress. Neuroreport 16:597-601 https://doi.org/10.1097/00001756-200504250-00016
  11. Korinek V(1998): Depletion of epithelial stem-cell compartments in the small intestine of mice lacking Tcf-4. Nature Genet 19:379-383 https://doi.org/10.1038/1270
  12. Lee BY, Kleber M, Hari L, Brault V, Suter U, Taketo MM, Kemler R, Sommer L (2004): Instructive role of Wnt/-catenin in sensory fate specification in neural crest stem cells. Science 303:1020-1023 https://doi.org/10.1126/science.1091611
  13. Lee JB, Lee JE, Park JH, Kim SJ, Kim MK, Roh SI, Yoon HS (2005): Establishment and maintenance of human embryonic stem cell lines on human feeder cells derived from uterine endometrium under serum-free condition. Biol Reprod 72:42-49 https://doi.org/10.1095/biolreprod.104.033480
  14. Loebel AF, Watson CW, Young AD, Tam PL (2003): Lineage choice and differentiation in mouse embryos and embryonic stem cells. Dev Biol 264:1-14 https://doi.org/10.1016/S0012-1606(03)00390-7
  15. Lynn A, Hanna RK, Foreman IA, Tarasenko DS, Kessler, Patricia AL (2002): Requirement for Foxd3 in maintaining pluripotent cells of the early mouse embryo. Genes Dev 16:2650-2661 https://doi.org/10.1101/gad.1020502
  16. Matsuda T, Nakamura T, Nakao K, Arai T, Katsuki M, Heike T, Yokota T (1999): STAT3 activation is sufficient to maintain an undifferentiated state of mouse embryonic stem cells. EMBO J 18:4261-4269 https://doi.org/10.1093/emboj/18.15.4261
  17. Meijer L, Flajolet M, Greengard P (2004): Pharmacological inhibitors of glycogen synthase kinase 3. Trends Pharmacol Sci 25:471-80 https://doi.org/10.1016/j.tips.2004.07.006
  18. Meijer L, Skaltsounis A, Magiatis P (2003): GSK-3-selective inhibitors derived from tyrian purple indirubins. Chem Biol 10:1255-1266 https://doi.org/10.1016/j.chembiol.2003.11.010
  19. Mitsui K, Tokuzawa Y, Itoh H, Segawa K, Murakami M, Takahashi K, Maruyama M, Maeda M, Yamanaka S (2003): The homeoprotein nanog is required for maintenance of pluripotency in mouse epiblast and ES cells. Cell 113:631-642 https://doi.org/10.1016/S0092-8674(03)00393-3
  20. Moon RT, Bowerman B, Boutros M, Perrimon N (2002): The promise and perils of Wnt signaling through $\beta$-catenin, Science 296:1644-1646 https://doi.org/10.1126/science.1071549
  21. Murdoch B, Chadwick K, Martin M, Shojaei F, Shah KV, Gallacher L, Moon RT, Bhatia M (2003): Wnt-5a augments repopulating capacity and primitive hema-topoietic development of human blood stem cells in vivo. Proc Nati Acad Sci USA 100:3422-3427 https://doi.org/10.1073/pnas.0130233100
  22. Niwa H, Burdon TM, Chambersm I, Smith A (1998): Self-renewal of pluripotent embryonic stem cells is mediated via activation of STAT3. Genes Dev 12:2048-2060 https://doi.org/10.1101/gad.12.13.2048
  23. Nichols J, Zevnik B, Anastassiadis K, Niwa H, Klewe-Nebenius D, Chambers I, Scholer H, Smith A (1998): Formation of pluripotent stem cells in the mammalian embryo depends on the POU transcription factor Oct4. Cell 95:379-391 https://doi.org/10.1016/S0092-8674(00)81769-9
  24. Park JH, Kim SJ, Oh EJ, Moon SY, Roh SI, Kim CG, Yoon HS (2003): Establishment and maintenance of human embryonic stem cells on STO, a permanently growing cell line. Biol Reprod 69:2007-2014 https://doi.org/10.1095/biolreprod.103.017467
  25. Polakis P (2002): Casein kinase 1: a Wnt'er of disconnect. Curr Biol 12:R499-R501 https://doi.org/10.1016/S0960-9822(02)00969-7
  26. Poluchronopoulos P, Magiatis P, Skaltsounis A (2004): Structural basis for the synthesis of indirubins as potent and selective inhibitors of glycogen synthase kinase-3 and eyelin-dependent kinases. J Med Chem 47:935-946 https://doi.org/10.1021/jm031016d
  27. Reya T (2003): A role of Wnt signaling in self-renewal of hematopoietic stem cells. Nature 423:409-414 https://doi.org/10.1038/nature01593
  28. Sato N, Meijer L, Skaltsounis L, Greengard P, Brivanlou AH (2004): Maintenance of pluripoteney in human and mouse embryonic stem cells through activation of Wnt signaling by a GSK-3-specific inhibitor. Nature Med 10:55-63 https://doi.org/10.1038/nm979
  29. Sato N, Sanjuan IM, Heke M, Uchida M, Naef F, Brivanlou AH (2003): Molecular signature of human embryonic stem cells and its comparison with the mouse. Dev Biol 260:404-413 https://doi.org/10.1016/S0012-1606(03)00256-2
  30. Smith AG (2001): Embryo-derived stem cells: of mice and men. Annu Rev Cell Dev Biol 17:435-462 https://doi.org/10.1146/annurev.cellbio.17.1.435
  31. Stewart CL, Kaspar P, Brunet LJ, Bhatt H, Gadi I, Kontgen F, Abbondanzo SJ (1992): Blastocyst implantation depends on maternal expression of Leukaemia inhibitory factor. Nature 359:76-79 https://doi.org/10.1038/359076a0
  32. Thomson JA, Itskovitz-Eldor J, Shapiro SS, Waknitz MQJ, Marshall VS, Jones JM (1998): Embryonic stemlines derived from human blastocysts. Science 282: 1145-1147 https://doi.org/10.1126/science.282.5391.1145
  33. Williams RL, Hilton DJ, Pease S, Willson TA, Stewart CL, Gearing DP, Wagner EF, Metcalf D, Nicola NA, Gough NM (1988): Myeloid leukaemia inhibitory factor maintains the developmental potential of embryonic stem cells. Nature. 336:684-687 https://doi.org/10.1038/336684a0
  34. Zhu AJ, Watt FM (1999): $\beta$-catenin signaling modulates proliferative potential of human epidermal keratinocytes independently of intercellular adhesion. Development 126:2285-2298