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Seoul, South Korea

Gupta M.K.,Bio Organ Research Center | Uhm S.J.,Bio Organ Research Center | Lee H.T.,Bio Organ Research Center
Fertility and Sterility

Objective: To investigate the effect of vitrification and beta-mercaptoethanol (β-ME) on reactive oxygen species (ROS) activity and in vitro development of oocytes vitrified before or after in vitro fertilization (IVF). Design: Randomized prospective study. Setting: University-based assisted reproductive technology laboratory. Animals(s): Abattoir-derived porcine ovaries. Interventions(s): Oocytes were vitrified either before or 4 hours after the end of IVF by solid surface vitrification (SSV) without centrifugation and/or delipation procedure. β-ME was used to inhibit ROS activity. Main Outcome Measures(s): Viability was evaluated by membrane integrity and esterase enzyme activity using fluorescein diacetate staining while ROS activity was assessed by 2′,7′-dichlorofluorescein assay. Result(s): Vitrification increased the ROS activity and decreased the viability and in vitro development of vitrified oocytes. Addition of β-ME to vitrification and culture medium partially annihilated the ROS activity but did not improve the viability of vitrified-warmed oocytes. Furthermore, β-ME had no effect on improving the fertilization ability of oocytes vitrified at metaphase II stage but significantly increased their ability to cleave. β-ME also increased the rate of cleavage and blastocyst formation ability of oocytes vitrified 4 hours after the end IVF. Conclusion(s): Vitrification increases ROS activity in oocytes that can be partially annihilated by β-ME to obtain enhanced embryonic development. © 2010 American Society for Reproductive Medicine. Source

Jung Y.H.,Bio Organ Research Center | Gupta M.K.,Bio Organ Research Center | Oh S.H.,Bio Organ Research Center | Uhm S.J.,Bio Organ Research Center | Lee H.T.,Bio Organ Research Center
Experimental Cell Research

This study evaluated the essentiality of glial cell line-derived neurotrophic factor (GDNF) for in vitro culture of established mouse multipotent adult germline stem (maGS) cell lines by culturing them in the presence of GDNF, leukemia inhibitory factor (LIF) or both. We show that, in the absence of LIF, GDNF slows the proliferation of maGS cells and result in smaller sized colonies without any change in distribution of cells to different cell-cycle stages, expression of pluripotency genes and in vitro differentiation potential. Furthermore, in the absence of LIF, GDNF increased the expression of male germ-line genes and repopulated the empty seminiferous tubule of W/Wv mutant mouse without the formation of teratoma. GDNF also altered the genomic imprinting of Igf2, Peg1, and H19 genes but had no effect on DNA methylation of Oct4, Nanog and Stra8 genes. However, these effects of GDNF were masked in the presence of LIF. GDNF also did not interfere with the multipotency of maGS cells if they are cultured in the presence of LIF. In conclusion, our results suggest that, in the absence of LIF, GDNF alters the growth characteristics of maGS cells and partially impart them some of the germline stem (GS) cell-like characteristics. © 2009 Elsevier Inc. All rights reserved. Source

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