DOI QR코드

DOI QR Code

Propagation of Human Embryonic Stem Cells on Human Amniotic Fluid Cells as Feeder Cells in Xeno-Free Culture Conditions

  • Jung, Juwon (Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University) ;
  • Baek, Jin Ah (Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University) ;
  • Seol, Hye Won (Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University) ;
  • Choi, Young Min (Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University)
  • 투고 : 2016.03.08
  • 심사 : 2016.03.19
  • 발행 : 2016.03.31

초록

Human embryonic stem cells (hESCs) have been routinely cultured on mouse embryonic fibroblast feeder layers with a medium containing animal materials. For clinical application of hESCs, animal-derived products from the animal feeder cells, animal substrates such as gelatin or Matrigel and animal serum are strictly to be eliminated in the culture system. In this study, we performed that SNUhES32 and H1 were cultured on human amniotic fluid cells (hAFCs) with KO-SR XenoFree and a humanized substrate. All of hESCs were relatively well propagated on hAFCs feeders with xeno-free conditions and they expressed pluripotent stem cell markers, alkaline phosphatase, SSEA-4, TRA1-60, TRA1-81, Oct-4, and Nanog like hESCs cultured on STO or human foreskin fibroblast feeders. In addition, we observed the expression of nonhuman N-glycolylneuraminic acid (Neu5GC) molecules by flow cytometry, which was xenotransplantation components of contamination in hESCs cultured on animal feeder conditions, was not detected in this xeno-free condition. In conclusion, SNUhES32 and H1 could be maintained on hAFCs for humanized culture conditions, therefore, we suggested that new xeno-free conditions for clinical grade hESCs culture will be useful data in future clinical studies.

키워드

참고문헌

  1. Bajek A, Olkowska J, Gurtowska N, Kloskowski T, Walentowicz- Sadlecka M, Sadlecki P, Grabiec M, Drewa T (2014) Human amniotic-fluid-derived stem cells: a unique source for regenerative medicine. Expert Opin Biol Ther 14:831-839. https://doi.org/10.1517/14712598.2014.898749
  2. Cobo F, Stacey GN, Hunt C, Cabrera C, Nieto A, Montes R, Cortes JL, Catalina P, Barnie A, Concha A (2005) Microbiological control in stem cell bank: approaches to standardisation. Appl Microbiol Biotechnol 68:456-466. https://doi.org/10.1007/s00253-005-0062-2
  3. Crook JM, Peura TT, Kravets L, Bosman AG, Buzzard JJ, Horne R, Hentze H, Dunn NR, Zweigerdt R, Chua F, Upshall A, Colman A (2007) The generation of six clinical-grade human embryonic stem cell lines. Cell Stem Cell 1:490-494. https://doi.org/10.1016/j.stem.2007.10.004
  4. Ding DC, Shyu WC, Lin SZ, Liu HW, Chiou SH, Chu TY (2012) Human umbilical cord mesenchymal stem cells support nontumorigenic expansion of human embryonic stem cells. Cell Transplant 21:1515-1527. https://doi.org/10.3727/096368912X647199
  5. Ellerstrom C, Strehl R, Moya K, Andersson K, Bergh C, Lundin K, Hyllner J, Semb H (2006) Derivation of a xeno-free human embryonic stem cell line. Stem Cells 24:2170-2176. https://doi.org/10.1634/stemcells.2006-0130
  6. Galende E, Karakikes I, Edelmann L, Desnick RJ, Kerenyi T, Khoueiry G, Lafferty J, McGinn JT, Brodman M, Fuster V, Hajjar RJ, Polgar K (2010) Amniotic fluid cells are more efficiently reprogrammed to pluripotency than adult cells. Cell Reprogram 12:117-125. https://doi.org/10.1089/cell.2009.0077
  7. Havasi P, Nabioni M, Soleimani M, Bakhshandeh B, Parivar K (2013) Mesenchymal stem cells as an appropriate feeder layer for prolonged in vitro culture of human induced pluripotent stem cells. Mol Biol Rep 40:3023-3031. https://doi.org/10.1007/s11033-012-2376-3
  8. Hovatta O (2006) Derivation of human embryonic stem cell lines, towards clinical quality. Reprod Fertil 18:823-828. https://doi.org/10.1071/RD06075
  9. Ilic D, Giritharan G, Zdravkovic T, Caceres E, Genbacev O, Fisher SJ, Krtolica A (2009) Derivation of human embryonic stem cell lines from biopsied blastomeres on human feeders with minimal exposure to xenomaterials. Stem Cells Dev 18:1343-1350. https://doi.org/10.1089/scd.2008.0416
  10. Ilic D, Stephenson E, Wood V, Jacquet L, Stevenson D, Petrova A, Kadeva N, Codognotto S, Patel H, Semple M, Cornwell G, Ogilvie C, Braude P (2012) Derivation and feeder-free propagation of human embryonic stem cells under xeno-free conditions. Cytotherapy 14:122-128. https://doi.org/10.3109/14653249.2011.623692
  11. Kim HS, Seol HW, Ahn HJ, Oh SK, Ku SY, Kim SH, Choi YM, Kim JG, Moon SY (2004) Human amniotic fluid cells support expansion culture of human embryonic stem cells. Kor J Fertil Steril 31:261-271.
  12. Kook M, Park S, Kang HM, Kim H (2006) In vitro differentiation of human amniotic membrane-derived stem cells into hepatocyte like cells. Dev Reprod 10:63-73.
  13. Ma X, Li H, Xin S, Ma Y, Ouyang T (2014) Human amniotic fluid stem cells support undifferentiated propagation and pluripotency of human embryonic stem cell without b-FGF in a density dependent manner. Int J Clin Exp Pathol 7:4661-4673.
  14. Mallon BS, Park KY, Chen KG, Hamilton RS, McKay RD (2006) Toward xeno-free culture of human embryonic stem cells. Int J Biochem Cell Biol 38:1063-1075. https://doi.org/10.1016/j.biocel.2005.12.014
  15. Martin MJ, Muotri A, Gage F, Varki A (2005) Human embryonic stem cells express an immunogenic nonhuman sialic acid. Nat Med 11:228-232. https://doi.org/10.1038/nm1181
  16. Park Y, Kim JH, Lee SJ, Choi IY, Park SJ, Yoo YD, Geum DH, Choi CW, Kim SH, Kim BS (2011) Human feeder cells can support the undifferentiated growth of human and mouse embryonic stem cells using their own basic fibroblast growth factors. Stem Cells Dev 20:1901-1910. https://doi.org/10.1089/scd.2010.0496
  17. Prusa AR, Hengstschlager M (2002) Amniotic fluid cells and human stem cell research: a new connection. Med Sci Monit 8:RA253-257.
  18. Prusa AR, Marton E, Rosner M, Bernaschek G, Hengstschlager M (2003) Oct-4 expressing cells in human amniotic fluid: a new source for stem cell research? Hum Reprod 18:1489-1493. https://doi.org/10.1093/humrep/deg279
  19. Richards M, Fong CY, Chan WK, Wong PC, Bonso A (2002) Human feeders support prolonged undifferentiated growth of human inner cell masses and embryonic stem cells. Nat Biotechnol 20:933-936. https://doi.org/10.1038/nbt726
  20. Skottman H, Narkilahti S, Hovatta O (2007) Challenges and approaches to the culture of pluripotent human embryonic stem cells. Regen Med 2:265-273. https://doi.org/10.2217/17460751.2.3.265
  21. Soong YK, Huang SY, Yeh CH, Wang TH, Chang KH, Cheng PJ, Shaw SW S (2013) The use of human amniotic fluid mesenchymal stem cells as the feeder layer to establish human embryonic stem cell lines. J Tissue Eng Regen Med DOI:10.1002term.1702. [Epub ahead of print]
  22. Stacey GN, Cobo F, Nieto A, Talavera P, Healy L, Concha A (2006) The development of 'feeder' cells for the preparation of clinical grade hES cell lines: challenges and solutions. J Biotechnol 125:583-588. https://doi.org/10.1016/j.jbiotec.2006.03.011
  23. Tannenbaum SE, Turetsky TT, Singer O, Aizenman E, Kirshberg S, Ilouz N, Gil Y, Berman-Zaken Y, Perlman TS, Geva N, Levy O, Arbell D, Simon A, Ben-Meir A, Shufaro Y, Laufer N, Reubinoff BE (2012) Derivation of xeno-free and GMP-grade human embryonic stem cells-platforms for future clinical applications. PLoS One 7:e35325. https://doi.org/10.1371/journal.pone.0035325
  24. Thomson JA, Itskovitz-Eldor J, Shapiro SS, Waknitz MA, Swiergiel JJ, Marshall VS, Jones JM (1998) Embryonic stem cell lines derived from human blastocysts. Science 282:1145-1147. https://doi.org/10.1126/science.282.5391.1145
  25. Villa-Diaz LG, Ross AM, Lahann J, Krebsbach PH (2013) Concise review: The evolution of human pluripotent stem cell culture: from feeder cells to synthetic coatings. Stem Cells 31:1-7. https://doi.org/10.1002/stem.1260
  26. Wang Y, Chou BK, Dowey S, He C, Gerecht S, Cheng L (2013) Scalable expansion of human induced pluripotent stem cells in the defined xeno-free E8 medium under adherent and suspension culture conditions. Stem Cell Res 11:1103-1116. https://doi.org/10.1016/j.scr.2013.07.011