Evaluation of propanediol, ethylene glycol, sucrose and antifreeze proteins on the survival of slow-cooled mouse pronuclear and 4-cell embryos. Embryonic stem cell lines from human blastocysts: somatic differentiation in vitro. Reubinoff BE, Pera MF, Fong CY, Trounson A and Bongso A. Effective cryopreservation of human embryonic stem cells by the open pulled straw vitrification method. Reubinoff BE, Pera MF, Vajta G and Trounson AO. Ice-free cryopreservation of mouse embryos at - 196'C by vitrification. Chromosome and spindle configurations of human oocytes matured in vitro after cryopreservation at the germinal vesicle stage. Park SE, Lee KA, Son WY, Ko JJ, Lee SH and Cha KY. Development into blastocysts of bovine oocyte cryopreserved by ultra-rapid cooling. Potential Importance of Vitrification in Reproductive. Liebermann J, Nawroth F, Isachenko V, Isachenko E, Rahimi G and Tucker MJ. Vitrification of mouse and human blastocysts using a novel cryoloop container-less technique. Adapting a proven method for flashcooling protein crystals to the cryopreservation of live cells. Lane M, Bavister BD, Lyons EA and Forest KT. Human embryonic stem cells: the future is now. Cryopreservation: Freezing and Vitrification. Improved human oocyte development after vitrification: a comparison of thawing methods. Hong SW, Chung HM, Lim JM, Ko JJ, Yoon TK, Yee Bill and Cha KY 1999. New potential for human embryonic stem cells. Establishment in culture of pluripotential cells from mouse embryos. Human Reprod., 15:905-910Įvans MJ and Kaufman MH. Comparison of ethylene glycol, 1,2-propanediol and glycerol for cryopreservation of slow cooled mouse zygotes, 4cell embryos and blastocysts. Steril., 73: 545-551Įmiliani S, Bergh MV, Vannin N-S, Biramane J and Englert Y. In vitro blastocyst formation of human oocytes obtained from unstimulated and stimulated cycles after vitrification at various maturational stages. Steril., 74: 838-839Ĭhung HM, Hong SW, Lim JM, Lee SH, Cha WT, Ko JJ, Han SY, Choi DH and Cha KY.
Pregnancy and delivery of healthy infants developed from vitrified blastocysts in an IVF-ET program. Reprod., 15:2598-2603Ĭhoi DH, Chung HM, Lim JM, Ko JJ, Yoon TK and Cha KY. Open pulled straws for vitrification of mature mouse oocytes preserve patterns of meiotic spindles and chromosomes better than conventional straws. Vitrification of embryos and oocytes with 5.5 mol/l ethylene glycol and 1.0 mol/l sucrose. Cell Endocrine, 169: 43-47Ĭhen SU, Ho HN and Yang YS. Fertil., 99:471-477Ĭha KY, Chung HM, Lim JM, Ko JJ, Han SY, Choi DH and Yoon TK. Design of vitrification solutions for the cryopreservation of embryos. These results show that vitrification method by using EM grid supplemented with EG and PROH in hES cells may be most efficient at present to minimize cyto-toxicity and cellular damage derived by ice crystal formation and furthermore may be employed for clinical application.Īli J and Shelton N.
Through RT-PCR analysis showed Oct-4 gene expression was down-regulated and embryonic germ layer markers were up-regulated in the vitrified hES cells during spontaneous differentiation. The vitrified hES cells retained the normal morphology, alkaline phosphates activity, and the expression of SSEA-3 and 4. The hES cells vitrified with EM grid showed relatively constant colony forming efficiency and survival rates, compared to those of unverified hES cells.
It shows high survival rates of hES cells frozen with EG and DMSO (60.8%), or EG and PROH(65.8%) on EM grids better than those of OPS, compared to those frozen with EG alone (2.4%) or PROH alone (0%) alone. Survival rates of vitrified hES cells were assessed by counting of undifferentiated colonies. Thawed hES cells were dipped into sequentially holding media with 0.2 M sucrose for 1 min, 0.1 M sucrose for 5 min and holding media for 5 min twice and plated onto a fresh feeder layer.
We compared to types of vehicles, such as open pulled straw (OPS) or electron microscopic cooper grids (EM grids). For the vitrifications, ES cells are exposed to the 4 different cryoprotectants, ethylene glycol (EG), 1,2-propanediol (PROH), EG with dime-thylsulfoxide (DMSO) and EG with PROH. The objective of this study is to improve freezing method with less cryo-injuries and best survival rates in hES cells by comparing various vitrification conditions. Human embryonic stem (hES) cell lines have been derived from human blastocysts and are expected to have far-reaching applications in regenerative medicine.