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Lee E.J.,Seoul National University | Lee H.-N.,Seoul National University | Kang H.-J.,Seoul National University | Kim K.-H.,Seoul National University | And 10 more authors.
Tissue Engineering - Part A | Year: 2010

Application of human embryonic stem cells (hESCs) to stem-cell therapy is not feasible because of the risk of tumorigenicity and rejection. In contrast, human mesenchymal stem cells (hMSCs) are free from the risk of tumorigenicity and also have immune privilege. However, hMSCs obtained from adults have infinite variety in terms of the biological characteristics and functionality. We report here a new derivation method of hMSCs from hESCs. The derivation of hMSCs from three different hESC lines (SNUhES3, CHA3-hESC, and H9) was performed by embryoid bodies formation and subsequent culture with stage-different media without using inductive xenogenic feeder and mechanical selection procedure. The derived cells were morphologically similar to the unique fingerprint-like pattern of hMSCs and grew stably for at least 35 passages in vitro. These cells had hMSCs-like immunophenotypes: negative for CD34 and CD45; positive for CD29, CD44, CD73, CD90, and CD105. They could be differentiated into multiple lineages including osteocytes, chondrocytes, adipocytes, and myocytes. They maintained normal karyotype during the long-term cultivation and did not show tumorigenicity when transplanted into the immunodeficient mice. In conclusion, the new embryoid body-based derivation method of hMSCs from hESCs is simple, safe, and reproducible in three different hESC lines. We expect that this method will provide a more effective and powerful tool to derive hMSCs from various hESC lines. Copyright 2010, Mary Ann Liebert, Inc. Source


Chae S.J.,Maria Fertility Hospital | Kim J.J.,Seoul National University | Kim J.J.,Healthcare System Gangnam Center | Choi Y.M.,Seoul National University | And 5 more authors.
Gynecologic and Obstetric Investigation | Year: 2010

Background/Aims: To investigate whether the peroxisome proliferator- activated receptor-γ (PPAR-γ) Pro12Ala and the PPAR-γ coactivator-1α (PGC-1α) Gly482Ser polymorphisms were associated with polycystic ovary syndrome (PCOS). Methods: Genetic analyses of the PPAR-γ Pro12Ala and the PGC-1α Gly482Ser polymorphisms were performed in 184 patients with PCOS and 256 controls. Hormone levels, biochemical and clinical features were analyzed according to these polymorphisms. Results: Neither the PPAR-γPro12Ala nor the PGC-1α Gly482Ser polymorphism showed significant differences in genotypic distribution between women with PCOS and controls. In PCOS patients, women with the non-Pro/Pro genotypes of the PPAR-γ Pro12Ala polymorphism showed statistically significantly higher HDL levels than those with the Pro/Pro genotype (p = 0.002). PCOS patients who had the Ser/Ser genotype of the PGC-1α Gly482Ser polymorphism had significantly higher levels of postprandial 2-hour insulin than those with the Gly/Ser genotype (p = 0.045). Conclusions: Neither the PPAR-γPro12Ala nor the PGC-1α Gly482Ser polymorphism were supposed to be susceptible genes in PCOS. However, in PCOS patients, the PPAR-γPro12Ala and the PGC-1α Gly482Ser polymorphism may modulate the concentrations of serum HDL levels and postprandial 2-hour insulin level, respectively. © 2010 S. Karger AG, Basel. Source


Park K.E.,Seoul National University | Ku S.-Y.,Seoul National University | Ku S.-Y.,Institute of Reproductive Medicine and Population | Jung K.C.,Seoul National University | And 10 more authors.
Reproductive Sciences | Year: 2013

Gonadotropins including follicle-stimulating hormone (FSH) and luteinizing hormone (LH) play a crucial role in human-assisted reproduction techniques. Despite wide use of recombinant gonadotropins in clinical practice, the efficacy of urinary gonadotropins and the dosage of LH component have not yet been elucidated. This study was designed to investigate the difference of follicle culture outcomes according to various compositions of gonadotropins during in vitro culture of mouse preantral follicles. Ovaries were obtained from the 14-day-old C57BL/6 mice, and preantral follicles were isolated and cultured in culture media supplemented with human menopausal gonadotropin (hMG) 200 mIU/mL (group 1), recombinant FSH and LH (rFSH + rLH) 200 mIU/mL each (group 2), rFSH 200 mIU/mL + rLH 100 mIU/mL (group 3), or rFSH 200 mIU/mL + rLH 20 mIU/mL (group 4). Follicle survival rate was significantly lower in group 4. Antral follicles in lower doses of LH (groups 3, 4) showed a statistically significant larger diameter and tended to have a higher antral formation rate. However, follicles in group 1 tended to have a higher oocyte maturation rate. Estradiol concentration from conditioned media from 2:1 FSH/LH (group 3) was significantly higher than those from 1:1 FSH/LH (group 2) or 10:1 FSH/LH (group 4). Half dose of rLH to rFSH facilitated upregulation of growth differentiation factor 9 (Gdf9) expression in granulosa cells when compared to 1:1 FSH/LH or 10:1 FSH/LH. Conclusively, recombinant gonadotropins provided a comparable condition to hMG, and half dose of rLH to rFSH seems to be more suitable for follicular development during in vitro culture. © 2012 The Author(s). Source


Lee J.R.,Seoul National University | Kim S.H.,Seoul National University | Kim S.H.,Institute of Reproductive Medicine and Population | Jee B.C.,Seoul National University | And 5 more authors.
Fertility and Sterility | Year: 2011

This study was performed to compare antimüllerian hormone (AMH) levels measured by two commercially available AMH measuring kits currently available, and to evaluate the AMH levels as predictor of controlled ovarian hyperstimulation (COH) outcome using the two different kits. The two assays for AMH measurement provide similar results, and serum AMH levels measured by the two kits both could be used as COH outcome predictors with similar reference values. © 2011 by American Society for Reproductive Medicine. Source


Kim Y.Y.,Institute of Reproductive Medicine and Population | Ku S.-Y.,Institute of Reproductive Medicine and Population | Ku S.-Y.,Seoul National University | Oh S.K.,Institute of Reproductive Medicine and Population | And 7 more authors.
Tissue Engineering and Regenerative Medicine | Year: 2011

Human embryonic stem cell (hESC)-derived, differentiated cells are an alternative cell source for many intractable diseases in regenerative medicine. However, the proper environments for long-term storage of hESCderived cells are still unknown. In this study, we compared the recovery rate of two different cryopreservation methods: mass cryopreservation vs. vitrification. hESC-derived cardiomyocytes (CMs) at two stages, day 13 (pre-contraction) and day 16 (post-contraction), were frozen in the presence of ROCK inhibitors. Thawed-CMs of day 13 from both groups showed higher survival rates than those of day 16. Thawed cells retained the characteristics of cardiomyocytes after long-term storage. Contraction of thawed CMs appeared earlier in vitrification group than in mass cryopreserved group. This study demonstrated that vitrification can be a useful method for the long-term storage of hESC-derived CMs. Source

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