Toxic Effect of Cryoprotectants on Embryo Development in a Murine Model / 대한불임학회지

OBJECTIVES: The aim of this study was to assess toxicities of cryoprotectants. METHODS: Toxicities of two cryoprotectants, dimethyl sulfoxide (DMSO) and 1,2-propanediol (PROH), were investigated using a murine embryo model. Female F-1 mice were stimulated with gonadotropin, induced ovulation with hCG and mated. Two cell embryos were collected and cultured after exposure to either DMSO or PROH. Embryo development was evaluated up to the blastocyst stage. Blastocysts were stained with bis-benzimide to evaluate the cell count and with terminal deoxynucleotidyl transferase mediated dUTP nick labeling (TUNEL) to assess apoptosis. RESULTS: The total cell count of blastocysts that were treated with DMSO at the 2-cell stage was significantly lower than that were treated with PROH (75.9+/-27.0) or the control (99.0+/-18.3) (p<0.001). On comparison of two cryoprotectant treated groups, the DMSO treated group showed a decreased cell count compared with the PROH treated group (p<0.05). Both DMSO (14.2+/-1.5) and PROH (11.2+/-1.4) treated groups showed higher apoptosis rates of cells in the blastocyst compared with the control (6.2+/-0.9, p<0.0001). In addition, the DMSO treated group showed more apoptotic cells than the PROH treated group (p<0.001). CONCLUSIONS: The potential toxicity of cryoprotectants was uncovered by prolonged exposure of murine embryos to either DMSO or PROH at room temperature. When comparing two cryoprotective agents, PROH appeared to be less toxic than DMSO at least in a murine embryo model.

Vitamin B Status and Serum Homocysteine Levels in Infertile Women

Adequate vitamin B(2), vitamin B(6), folate and vitamin B(12) nutrition is known to be important for reproductive function in women of childbearing age. The purpose of this study was to evaluate serum vitamin B(2), vitamin B(6), folate and vitamin B(12) status and serum homocysteine levels in 115 women aged 33.2 +/- 4.0 years, who had been diagnosed with infertility, and 49 women aged 34.5 +/- 3.8 years having at least one born child. Total vitamin B(2) and vitamin B(6) intakes in infertile women were significantly lower than those in control. Serum vitamin B(2), vitamin B(6), folate and vitamin B(12) concentrations were significantly lower in infertile women than those in control and serum homocysteine levels were significantly higher in infertile women than those in control. Thirteen percent in infertile women and zero percent in control were assessed as hyperhomocysteinemic and there was a significant difference in the prevalence of hyperhomocysteinemia between infertile women and control. 41% infertile women were assessed as folate deficiency. Serum folate concentrations was negatively correlated with serum homocysteine of the infertile women and control. Total vitamin B2 intakes was negatively correlated with serum homocysteine of the infertile women and control. Total vitamin B6, folate intakes were negatively correlated with homocysteine of infertile women only. In conclusion, infertile women are needed to intake more B vitamins intakes. Furthermore researches are needed to estimate adequate B vitamin supplementation in infertile women.