Efficient derivation of embryonic stem cells and primordial germ cell-like cells in cattle.

The induction of the germ cell lineage from pluripotent stem cells (in vitro gametogenesis) will help understand the mechanisms underlying germ cell differentiation and provide an alternative source of gametes for reproduction. This technology is especially important for cattle, which are among the most important livestock species for milk and meat production. Here, we developed a new method for robust induction of primordial germ cell-like cells (PGCLCs) from newly established bovine embryonic stem (bES) cells. First, we refined the pluripotent culture conditions for pre-implantation embryos and ES cells. Inhibition of RHO increased the number of epiblast cells in the pre-implantation embryos and dramatically improved the efficiency of ES cell establishment. We then determined suitable culture conditions for PGCLC differentiation using bES cells harboring BLIMP1-tdTomato and TFAP2C-mNeonGreen (BTTN) reporter constructs. After a 24-h culture with bone morphogenetic protein 4 (BMP4), followed by three-dimensional culture with BMP4 and a chemical agonist and WNT signaling chemical antagonist, bES cells became positive for the reporters. A set of primordial germ cells (PGC) marker genes, including PRDM1/BLIMP1, TFAP2C, SOX17, and NANOS3, were expressed in BTTN-positive cells. These bovine PGCLCs (bPGCLCs) were isolated as KIT/CD117-positive and CD44-negative cell populations. We anticipate that this method for the efficient establishment of bES cells and induction of PGCLCs will be useful for stem cell-based reproductive technologies in cattle.

Induction of primordial germ cell-like cells from common marmoset embryonic stem cells by inhibition of WNT and retinoic acid signaling.

Reconstitution of the germ cell lineage using pluripotent stem cells provides a unique platform to deepen our understanding of the mechanisms underlying germ cell development and to produce functional gametes for reproduction. This study aimed to establish a culture system that induces a robust number of primordial germ cell-like cells (PGCLCs) from common marmoset (Callithrix jacchus) embryonic stem cells. The robust induction was achieved by not only activation of the conserved PGC-inducing signals, WNT and BMP4, but also temporal inhibitions of WNT and retinoic acid signals, which prevent mesodermal and neural differentiation, respectively, during PGCLC differentiation. Many of the gene expression and differentiation properties of common marmoset PGCLCs were similar to those of human PGCLCs, making this culture system a reliable and useful primate model. Finally, we identified PDPN and KIT as surface marker proteins by which PGCLCs can be isolated from embryonic stem cells without genetic manipulation. This study will expand the opportunities for research on germ cell development and production of functional gametes to the common marmoset.

Diurnal rhythms in fetal heart rate baseline and sustained fetal tachycardia in twin pregnancy.

OBJECTIVE: To elucidate the synchronization of phases of diurnal rhythms in fetal heart rate (FHR) baseline between twin fetuses and the occurrence of sustained fetal tachycardia. METHODS: FHR was simultaneously recorded in twins for 24 h in 7 monochorionic diamniotic (MD) and 8 dichorionic diamniotic (DD) twin pregnancies at 35 to 38 weeks of gestation. The diurnal rhythms of the hourly mean FHR baseline were tested in each fetus, and the time of occurrence of sustained fetal tachycardia was compared between twins. The correlation coefficients and phase lags of diurnal rhythms between the hourly mean FHR baselines of twins were calculated in each case. RESULTS: There were significant diurnal rhythms in the hourly mean FHR baselines of all twin fetuses (p<0.01). The patterns of diurnal rhythms were similar for each pair of twins, with the exception of the periods of sustained fetal tachycardia. Sustained fetal tachycardia was not coincident between twins. Analysis in which the periods of sustained fetal tachycardia were excluded demonstrated a significant correlation between the hourly mean FHR baselines of twins in all cases (p<0.01). In the case of DD twins, the phase lag between twins was 0; however, phase lags were observed in 4 cases of MD twins. CONCLUSIONS: The results of this study revealed that the diurnal rhythms in the FHR baseline correlated well between twins, and that the occurrences of sustained fetal tachycardia were completely independent.

Analysis of the influence of diurnal variation in maternal movements on fetal heart rate acceleration.

To determine the influence of diurnal variation in maternal movements (MM) on fetal heart rate (FHR) accelerations, 24-hour maternal heart rates (MHR), MM and FHR recordings were made simultaneously on nine healthy pregnant women at 36-39 weeks of gestation. Correlations between the diurnal variations in maternal factors and FHR parameters; that is, baseline FHR, amplitude, duration and hourly percentage of time spent in accelerations, were examined. Only a few cases showed a significant correlation between acceleration parameters and the diurnal variation of MHR or MM, while most cases indicated a high correlation between the diurnal variation in baseline FHR and that of maternal factors. In conclusion, the occurrence and form of accelerations might be independent of maternal activity.

Differences in fractal structure of adult heart rate fluctuation between successive rapid eye movement sleeps: case report.

The dynamics of fractal structures of heart rate (HR) fluctuations in successive rapid eye movement sleeps (REMS) was investigated in a healthy male. Fractal analysis using a purposely-devised method was carried out at 10-min time series of HR, which were being recorded simultaneously by an electroencephalogram at night. Scaling relationships showed three slopes (alpha(s)). The mean alphas below one cross-over point (< 6.0 s), mean alpha(m) between two cross-over points, and mean alpha1 above another cross-over point (> 12.0 s) during REMS changed from 2.0, 1.0, and 0.3 in the third cycle to 2.0, 0.9, and 1.2 in the fourth cycle. The present study demonstrates the differences in multiscaled fractal structures of HR fluctuations between the successive REMS in a particular case.