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OBJECTIVE: To determine whether animal age impacts in vitro preantral follicle growth. Effects of hCG, stem cell factor (SCF), and/or insulin-like growth factor (IGF) supplementation in growth medium were also investigated. METHODS: Intact preantral follicles were mechanically isolated from fresh ovaries of BDF1 mice and cultured in growth medium for 9 to 11 days. Surviving follicles with antrum formation were transferred to maturation medium for 14 to 18 hours. Follicle survival, antrum formation, and retrieval of metaphase II (MII) oocytes were compared among three age categories (4-5, 7-8, and 10-11 week-old). By using 7- to 8-week-old mice, preantral follicles were cultured in growth medium supplemented with hCG (0, 5, or 10 mIU/mL), SCF (50 ng/mL), IGF-1 (50 ng/mL), and SCF+IGF-1. RESULTS: Seven- to eight-week-old mice showed a higher follicle survival and antrum formation and produced more MII oocytes compared to other groups. In the 7- to 8-week-old mice, supplementation of 5 mIU/mL hCG significantly enhanced the antrum formation but the percentage of MII oocytes was similar to that of the control. Supplementation of SCF+IGF-1 did not enhance follicle survival or antrum formation but the percentage of MII oocytes increased modestly (39.1%) than in the control (28.6%, statistically not significant). CONCLUSION: Seven- to eight-week-old mice showed better outcomes in growth of preantral follicles in vitro than 4- to 5- or 10- to 11-week-old mice. Supplementation of hCG enhanced antrum formation and supplementation of SCF+IGF-1 yielded more mature oocytes; hence, these should be considered in the growth of preantral follicles in vitro.
Animals ; Female ; Humans ; Insulin-Like Growth Factor I ; Metaphase ; Mice ; Oocytes ; Ovarian Follicle ; Ovary ; Stem Cell Factor ; Stem Cells

OBJECTIVE: The aim of this study is to compare the efficiency of a method for the cryopreservation of mouse blastocyst. METHODS: Mouse embryos were obtained at 2-cell stage and cultured to blastocyst stage in T6 medium supplemented with 10% fetal bovine serum. Morphologically normal blastocysts were collected and randomly divided to one control and four experimental groups. In control group, blastocysts were cultured in vitro continuously for additional two days. In group 2, blastocysts were exposed to vitrification solution (ethylene glycol) only without cryopreservation (exposure only group). In group 3, 4 and 5, blastocysts were cryopreserved by slow-freezing procedure with glycerol (slow-freezing group) or by vitrification procedure using EM grids (EM grids group) and cryoloop (cryoloop group), respectively. Frozen blastocysts were thawed and cultured for additional two days. Twenty four hours after thawing, some blastocysts were fixed and stained with Hoechst 33342 (bisbenzimide) and the number of nuclei in each blastocysts were counted to confirm the survival of blastocysts in experimental groups. RESULTS: Survival rate and hatching rate of the blastocysts in slow-freezing group (24 h: 72.4% and 66.0%, 48 h: 63.2% and 64.6%) and EM grids group (24 h: survival rate 77.3%, 48 h: 70.1% and 71.4%) were significantly lower (X2-test p<0.05) than those of control group (24 h: 93.4% and 86.0%, 48 h: 88.5% and 90.7%). In contrast, the survival rate and hatching rate of the blastocysts in cryoloop group (24 h: 84.1% and 84.1%, 48 h 79.3% and 87.7%) is well compared with those in the control group. The mean (+/-SD) cell number of blastocyst in the exposure only (89.2+/-11.5), EM grids (85.0+/-10.3) and cryoloop (89.0+/-11.0) groups, except slow-freezing group (79.0+/-10.0), were not significantly different from that of control group (93.1+/-13.9) 24 h after thawing (Student's t-test). CONCLUSION: This study demonstrates that higher survival rate of vitrified-thawed mouse blastocyst can be obtained using cryoloop as the embryo container at freezing rather than slow-freezing or vitrification using EM grids. The results of this study suggest that vitrification using cryoloop (with ethylene glycol) may be a preferable procedure for mouse blastocyst cryopreservation and could be applied to the human blastocyst cryopreservation.
Animals ; Blastocyst* ; Cell Count ; Cryopreservation ; Embryonic Structures ; Freezing ; Glycerol ; Humans ; Mice* ; Survival Rate ; Vitrification*

OBJECTIVES: The bone mineral density (BMD) of young aged women was not fully investigated because of the limited indication of BMD test. This study was to investigate mean Z-score of BMD in healthy young aged Korean women and to find out factors influencing the BMD score in her twenties and thirties. METHODS: A total 876 young aged (20-40 years) women who had taken dual energy X-ray absorptiometry (DXA) for BMD screening from 2010 to 2014 were analyzed retrospectively using demographic data including body mass index (BMI), waist circumference (WC), thyroid stimulating hormone (TSH), and serum lipid profile. We compared the two age groups, in her twenties and thirties, regarding BMD and other parameters.
Absorptiometry, Photon ; Body Mass Index ; Bone Density ; Female ; Hip ; Humans ; Mass Screening ; Retrospective Studies ; Spine ; Thyroid Gland ; Thyrotropin ; Waist Circumference