Purpose The aim of this study was to establish a simple tool to predict good-quality embryos in in vitro fertilization (IVF) by using cumulus cells (CCs) or peripheral blood cells (PBCs). Methods Mitochondrial DNA was extracted from CCs and PBCs in patients undergoing IVF. Using real-time polymerase chain reaction, mtDNA copy number in a single cell was calculated. Embryo quality was assessed when it was transferred or frozen. Results CCs were obtained from 60 oocyte cumulus-cell complexes (OCCCs) in 30 women, and PBCs were collected from 18 women. For the 30 women in the study, the median age was 37 years old (range, 24-43), and the mean body mass index was 21.4 (standard error, 2.0). mtDNA content of CCs and PBCs was highly correlated (Pearson's r = 0.900, p < 0.0001). The median mtDNA content of CCs for goodand poor-quality embryos was 140 and 57, respectively (p < 0.0001). The median mtDNA content of PBCs for goodand poor-quality embryos was 36 and 13, respectively (p = 0.604). The logistic regression model indicated that mtDNA content in CCs was the only parameter that predicted good-quality embryos (p = 0.020). The receiver operating characteristic curve for obtaining good-quality embryos by mtDNA copy number in CCs had an area under the curve of 0.823, and using a threshold of 86, positive and negative predictive values were 84.4 and 82.1 %, respectively. Conclusions The determination of mtDNA content in CCs can be used to predict good-quality embryos.

Using the novel method, manually counting the number of motile spermatozoa becomes unnecessary. The novel method presented here will increase the objectivity and convenience of using the SIT as a clinical indicator.

The voltammetric behavior of barium and lithium ions in acetonitrile (AN) was studied at polymer membrane ion-selective electrodes which showed Nernstian responses to those metal ions in AN, in order to elucidate the mechanism of their potentiometric responses. The membrane electrodes consisted of polyacrylamide coupled to such poly(oxyethylene) derivatives as tetraethylene glycol monododecyl ether, dibenzo-18-crown-6, and cryptand222B and coated on platinum or gold disk. The cyclic voltammograms for barium and lithium ions, recorded repeatedly after setting the electrodes at the rest potentials, showed a cathodic peak at −1.5±0.2 V vs. Ag/0.01 M Ag+ (AN). The cathodic peak current (icp) decreased markedly between the first and second scans, but reached a steady value after the third scan. The icp values after the third scan were diffusion controlled and considered to be due to a transfer of the metal ions from the solution into the membrane. The differences in icp between the first and the third scans were proportional to the potential scan rate and attributed to a transfer of the metal ions which had been accumulated at or near the surface of the membrane by combining with the functional groups of the polymer. The accumulation occurred rather slowly and saturated with time. Based on the fact that the response processes of the ion-selective electrodes were very much similar to the accumulation processes, the response mechanism of the ion-selective electrodes is discussed.

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Purpose In a previous study, a new method was described using the sperm immobilization test (SIT) with computer‐aided sperm analysis (CASA). However, obtaining high‐quality sperm as needed was a known issue. Here, we compared the results of using frozen‐thawed sperm and fresh sperm for the SIT using the CASA method. Methods For the frozen‐thawed preparation, 500 μL of condensed semen and 500 μL of Sperm Freeze were mixed in a cryovial and cryopreserved in liquid nitrogen. Density gradient centrifugation was used for the collection of motile sperm in both the fresh and frozen‐thawed sperm preparations. A total of 50 serum samples were prepared for both the fresh and frozen‐thawed sperm with each sample tested containing 10 μL of serum, 1 μL of either fresh or frozen motile sperm suspension, and 2 μL of complement. Sperm motilities were measured using CASA after a 1‐hour incubation period for both fresh and frozen‐thawed sperm. Results Both fresh and frozen‐thawed sperm reacted similarly when exposed to serum containing sperm‐immobilizing antibodies asserting the use of frozen‐thawed sperm for the diagnosis of immunological infertility. Conclusion These results suggest the possibility of using cryopreserved sperm for the SIT when fresh sperm is unavailable.