한국발생생물학회지:발생과생식 (Development and Reproduction)

  • 제14권3호
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  • Pages.155-162
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  • 2010
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  • 2465-9525(pISSN)
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  • 2465-9541(eISSN)
한국발생생물학회 (The Korean Society of Developmental Biology)
권호 표지

Isolation and Characterization of Trophoblast Stem Cells-like Cells Derived from Human Term Placenta

  • Na, Kyu-Hwan (Dept. of Biomedical Science, CHA University) ;
  • Shin, Kyung-Seon (Dept. of Biomedical Science, CHA University) ;
  • Choi, Jong-Ho (Dept. of Biomedical Science, CHA University) ;
  • Cha, Dong-Hyun (Dept. of Obstetrics and Gynecology, Kangnam CHA Medical Center, CHA University) ;
  • Kim, Gi-Jin (Dept. of Biomedical Science, CHA University)
  • 투고 : 2010.07.05
  • 심사 : 2010.08.03
  • 발행 : 2010.09.30
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초록

The trophectoderm is one of the earliest cell types to differentiate in the forming placenta. It is an important for the initial implantation and placentation during pregnancy. Trophoblast stem cells (TBSCs) develop from the blastocyst and are maintained by signals emanating from the inner cell mass. However, several limitations including rarity and difficulty in isolation of trophoblast stem cells derived from blastocyst still exist. To establish a model for trophoblast differentiation, we isolated TBSCs from human term placenta ($\geq$38 weeks) and characterized. Cell cycle was analyzed by measuring DNA content by FACS analysis and phenotype of TBSCs was characterized by RT-PCR and FACS analysis. TBSCs have expressed various markers such as self-renewal markers (Nanog, Sox2), three germ layer markers (hNF68, alpha-cardiac actin, hAFP), trophoblast specific markers (CDX-2, CK7, HLA-G), and TERT gene. In FACS analysis, TBSCs isolated from term placenta showed that the majority of cells expressed CD13, CD44, CD90, CD95, CD105, HLA-ABC, cytokeratin 7, and HLA-G. Testing for CD31, CD34, CD45, CD71, vimentin and HLA-DR were negative. TBSCs were shown to decrease the growth rate when cultured in conditioned medium without FGF4/heparin as well as the morphology was changed to a characteristic giant cell with a large cytoplasm and nucleus. In invasion assay, TBSCs isolated from term placenta showed invasion activities in in vivo using nude mice and in vitro Matrigel system. Taken together, these results support that an isolation potential of TBSCs from term placenta as well as a good source for understanding of the infertility mechanism.

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참고문헌

  1. Aplin JD, Straszewski-Chavez SL, Kalionis B, Dunk C, Morrish D, Forbes K, Baczyk D, Rote N, Malassine A, Knofler M (2006) Trophoblast differentiation: progenitor cells, fusion and migration - a workshop report. Placenta 27 Suppl A:S141-143. https://doi.org/10.1016/j.placenta.2006.01.011
  2. Benirshke K, Kaufmann P (2000) Pathology of the Human Placenta. New York: Springer Verlag.
  3. Blomberg L, Hashizume K, Viebahn C (2008) Blastocyst elongation, trophoblastic differentiation, and embryonic pattern formation. Reproduction 135:181-195. https://doi.org/10.1530/REP-07-0355
  4. Craven CM, Morgan T, Ward K (1998) Decidual spiral artery remodelling begins before cellular interaction with cytotrophoblasts. Placenta 19:241-252. https://doi.org/10.1016/S0143-4004(98)90055-8
  5. Douglas GC, VandeVoort CA, Kumar P, Chang TC, Golos TG (2009) Trophoblast stem cells: models for investigating trophectoderm differentiation and placental development. Endocr Rev 30:228-240. https://doi.org/10.1210/er.2009-0001
  6. Gerami-Naini B, Dovzhenko OV, Durning M, Wegner FH, Thomson JA, Golos TG (2004) Trophoblast differentiation in embryoid bodies derived from human embryonic stem cells. Endocrinology 145:1517-1524. https://doi.org/10.1210/en.2003-1241
  7. Golos TG, Pollastrini LM, Gerami-Naini B (2006) Human embryonic stem cells as a model for trophoblast differentiation. Semin Reprod Med 24:314-321. https://doi.org/10.1055/s-2006-952154
  8. Gude NM, Roberts CT, Kalionis B, King RG (2004) Growth and function of the normal human placenta. Thromb Res 114:397-407. https://doi.org/10.1016/j.thromres.2004.06.038
  9. Horne AW, White JO, Lalani EN (2000) The endometrium and embryo implantation. A receptive endometrium depends on more than hormonal influences. BMJ 321:1301-1302. https://doi.org/10.1136/bmj.321.7272.1301
  10. King A, Thomas L, Bischof P (2000) Cell culture models of trophoblast II: trophoblast cell lines - a workshop report. Placenta 21 Suppl A:S113-119. https://doi.org/10.1053/plac.1999.0526
  11. Knofler M, Simmons DG, Lash GE, Harris LK, Armant DR (2008) Regulation of trophoblast invasion - a workshop report. Placenta 29 Suppl A:S26-28. https://doi.org/10.1016/j.placenta.2007.11.008
  12. Lyall F (2005) Priming and remodelling of human placental bed spiral arteries during pregnancy - a review. Placenta 26 Suppl A:S31-36. https://doi.org/10.1016/j.placenta.2005.02.010
  13. Moore KL, Persaud TVN (2008) The Developing Human: Clinically Oriented Embryology. Philadelphia: Saunders Elsevier.
  14. Oda M, Shiota K, Tanaka S (2006) Trophoblast stem cells. Methods Enzymol 419:387-400. https://doi.org/10.1016/S0076-6879(06)19015-1
  15. Peiffer I, Belhomme D, Barbet R, Haydont V, Zhou YP, Fortunel NO, Li M, Hatzfeld A, Fabiani JN, Hatzfeld JA (2007) Simultaneous differentiation of endothelial and trophoblastic cells derived from human embryonic stem cells. Stem Cells Dev 16:393-402. https://doi.org/10.1089/scd.2006.0013
  16. Rossant J (2007) Stem cells and lineage development in the mammalian blastocyst. Reprod Fertil Dev 19:111- 118. https://doi.org/10.1071/RD06125
  17. Schenke-Layland K, Angelis E, Rhodes KE, Heydarkhan- Hagvall S, Mikkola HK, Maclellan WR (2007) Collagen IV induces trophoectoderm differentiation of mouse embryonic stem cells. Stem Cells 25:1529-1538. https://doi.org/10.1634/stemcells.2006-0729
  18. Schulz LC, Ezashi T, Das P, Westfall SD, Livingston KA, Roberts RM (2008) Human embryonic stem cells as models for trophoblast differentiation. Placenta 29 Suppl A:S10-16. https://doi.org/10.1016/j.placenta.2007.10.009
  19. Schumacher A, Brachwitz N, Sohr S, Engeland K, Langwisch S, Dolaptchieva M, Alexander T, Taran A, Malfertheiner SF, Costa SD, Zimmermann G, Nitschke C, Volk HD, Alexander H, Gunzer M, Zenclussen AC (2009) Human chorionic gonadotropin attracts regulatory T cells into the fetal-maternal interface during early human pregnancy. J Immunol 182:5488-5497. https://doi.org/10.4049/jimmunol.0803177
  20. Spitalieri P, Cortese G, Pietropolli A, Filareto A, Dolci S, Klinger FG, Giardina E, Di Cesare S, Bernardini L, Lauro D, Scaldaferri ML, Citro G, Novelli G, De Felici M, Sangiuolo F (2009) Identification of multipotent cytotrophoblast cells from human first trimester chorionic villi. Cloning Stem Cells 11:535-556. https://doi.org/10.1089/clo.2009.0046
  21. Xu RH, Chen X, Li DS, Li R, Addicks GC, Glennon C, Zwaka TP, Thomson JA (2002) BMP4 initiates human embryonic stem cell differentiation to trophoblast. Nat Biotechnol 20:1261-1264. https://doi.org/10.1038/nbt761