Fertility and Sterility
Volume 93, Issue 3 , Pages 976-985 , February 2010

High concentration of synthetic serum, stepwise equilibration and slow cooling as an efficient technique for large-scale cryopreservation of human embryonic stem cells

  • Ji Yeon Lee, M.Sc.

      Affiliations

    • Fertility Center, CHA General Hospital, CHA Research Institute, Pochon CHA University, Seoul, South Korea
    • ,
  • Jeoung Eun Lee, M.Sc.

      Affiliations

    • CHA Stem Cell Institute, Pochon CHA University, Seoul, South Korea
    • ,
  • Dong Ku Kim, Ph.D.

      Affiliations

    • CHA Stem Cell Institute, Pochon CHA University, Seoul, South Korea
    • ,
  • Tae Ki Yoon, M.D.

      Affiliations

    • Fertility Center, CHA General Hospital, CHA Research Institute, Pochon CHA University, Seoul, South Korea
    • ,
  • Hyung Min Chung, Ph.D.

      Affiliations

    • Fertility Center, CHA General Hospital, CHA Research Institute, Pochon CHA University, Seoul, South Korea
    • CHA Stem Cell Institute, Pochon CHA University, Seoul, South Korea
    • ,
  • Dong Ryul Lee, Ph.D.

      Affiliations

    • Fertility Center, CHA General Hospital, CHA Research Institute, Pochon CHA University, Seoul, South Korea
    • CHA Stem Cell Institute, Pochon CHA University, Seoul, South Korea
    • Corresponding Author InformationReprint requests: Dong Ryul Lee, Ph.D., Fertility Center, CHA General Hospital, CHA Research Institute, Pochon CHA University, 606-5 Yeoksam-dong, Gangnam-gu, Seoul 135–081, South Korea (FAX: 82-2-501-8704).

Received 27 August 2008 ,Revised 29 September 2008 ,Accepted 7 October 2008.

References 

  1. Thomson JA, Itskovitz-Eldor J, Shapiro SS, Waknitz MA, Swierqiel JJ, Marshall VS, et al. Embryonic stem cell lines derived from human blastocysts. Science. 1998;282:1145–1147
  2. Reubinoff BE, Pera MF, Fong CY, Trounson A, Bongso A. Embyonic stem cell lines from human blastocycts: somatic differentiation in vitro. Nat Biotechnol. 2000;8:399–404
  3. Reubinoff BE, Pera MF, Vajta G, Trounson A. Effective cryopreservation of human embryonic stem cells by the open pulled straw vitrification method. Hum Reprod. 2001;10:2187–2194
  4. Richards M, Fong CY, Tan S, Chan WK, Bongso A. An efficient and safe xeno-free cryopreservation method for the storage of human embryonic stem cells. Stem Cells. 2004;22:779–789
  5. Woods EJ, Liu J, Pollok K, Hartwell J, Smith FO, Williams DA, et al. A theoretically optimized method for cord blood stem cell cryopreservation. J Hematother Stem Cell Res. 2003;12:341–350
  6. Frederickx V, Michiels A, Goossens E, De Block G, Van Steirteghem AC, Tournaye H. Recovery, survival and functional evaluation by transplantation of frozen thawed mouse germ cells. Hum Reprod. 2004;19:948–953
  7. Ware CB, Nelson AM, Blau CA. Controlled-rate freezing of human ES cells. Biotechniques. 2005;38:879–883
  8. Yang PF, Hua TC, Wu J, Chang ZH, Tsung HC, Cao YL. Cryopreservation of human embryonic stem cells: a protocol by programmed cooling. Cryo Lett. 2006;27:361–368
  9. Ha SY, Jee BC, Suh CS, Kim HS, Oh SK, Kim SH, et al. Cryopreservation of human embryonic stem cells without the use of a programmable freezer. Hum Reprod. 2005;20:1779–1785
  10. Thonon F, Lens H, Touati K, Ector FJ, Delval A, Beckers JF, et al. A step-wise procedure for cryoprotectant equilibration improves the survival rate after thawing of in vitro produce embryos. [abstract] Theriogenology. 1995;43:338
  11. Ahn SE, Kim S, Park KH, Moon SH, Lee HJ, Kim GJ, et al. Primary bone-derived cells induce osteogenic differentiation without exogenous factors in human embryonic stem cells. Biochem Biophys Res Commun. 2006;340:403–408
  12. Cho SW, Moon SH, Lee SH, Kang SW, Kim J, Lim JM, et al. Improvement of postnatal neovascularization by human embryonic stem cell derived endothelial-like cell transplantation in a mouse model of hindlimb ischemia. Circulation. 2007;116:2409–2419
  13. Kim S, Kim GJ, Miyoshi H, Moon SH, Ahn SE, Lee JH, et al. Efficiency of the elongation factor-1alpha promoter in mammalian embryonic stem cells using lentiviral gene delivery systems. Stem Cells Dev. 2007;16:537–545
  14. Li T, Zhou C, Liu C, Mai Q, Zhuang G. Bulk vitrification of human embryonic stem cells. Hum Reprod. 2008;23:358–364
  15. Eum JH, Park JK, Lee WS, Cha KR, Yoon TK, Lee DR. Long-term liquid nitrogen vapor storage of mouse embryos cryopreserved using vitrification or slow cooling. Fertil Steril. Published online 29 April 2008 [Epub ahead of print].
  16. Katkov , Kim MS, Bajpai R, Altman YS, Mercola M, Loring JF, et al. Cryopreservation by slow cooling with DMSO diminished production of Oct-4 pluripotency market in human embryonic stem cells. Cryobiology. 2006;53:194–205

 J.Y.L. has nothing to disclose. J.E.L. has nothing to disclose. D.K.K. has nothing to disclose. T.K.Y. has nothing to disclose. H.M.C. has nothing to disclose. D.R.L. has nothing to disclose.

 Supported by a grant (SC1140) from the Stem Cell Research Center, funded by the Ministry of Education, Science and Technology, Korea.

PII: S0015-0282(08)04202-7

doi: 10.1016/j.fertnstert.2008.10.017

Fertility and Sterility
Volume 93, Issue 3 , Pages 976-985 , February 2010

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