Human blastocyst-derived, pluripotent cell lines are described that have normal karyotypes, express high levels of telomerase activity, and express cell surface markers that characterize primate embryonic stem cells but do not characterize other early lineages. After undifferentiated proliferation in vitro for 4 to 5 months, these cells still maintained the developmental potential to form trophoblast and derivatives of all three embryonic germ layers, including gut epithelium (endoderm); cartilage, bone, smooth muscle, and striated muscle (mesoderm); and neural epithelium, embryonic ganglia, and stratified squamous epithelium (ectoderm). These cell lines should be useful in human developmental biology, drug discovery, and transplantation medicine.

Embryonic Stem Cell Lines Derived from Human Blastocysts

OBJECTIVES: This report presents 2002 data on U.S. births according to a wide variety of characteristics. Data are presented for maternal demographic characteristics including age, live-birth order, race, Hispanic origin, marital status, and educational attainment; maternal characteristics (medical risk factors, weight gain, tobacco, and alcohol use); medical care utilization by pregnant women (prenatal care, obstetric procedures, complications of labor and/or delivery, attendant at birth, and method of delivery); and infant characteristics (period of gestation, birthweight, Apgar score, abnormal conditions, congenital anomalies, and multiple births). Also presented are birth and fertility rates by age, live-birth order, race, Hispanic origin, and marital status. Selected data by mother's State of residence are shown, as well as data on month and day of birth, sex ratio, and age of father. Trends in fertility patterns and maternal and infant characteristics are described and interpreted.

METHODS: Descriptive tabulations of data reported on the birth certificates of the 4.022 million births that occurred in 2002 are presented. Denominators for population-based rates are derived from the 2000 U.S. census. Rates for 1991-2001 may differ from those published previously based on the 1990 U.S. census.

RESULTS: There were 4,021,726 live births in 2002, essentially unchanged from 2001. The birth rate, fertility rate, and total fertility rates all declined 1 percent in 2002. The teenage birth rate dropped 5 percent, reaching another record low. The birth rates for women 20-24 years declined, whereas the rate for women 25-29 years was stable. The birth rate for women 30-34 years declined, but the rate for women 35-44 years continued to rise. Births to unmarried women changed very little. Smoking during pregnancy was down again. The timeliness of prenatal care continued to improve. The cesarean delivery rate climbed to the highest level ever reported in the United States (26.1 percent) and the rate of vaginal birth after previous cesarean plummeted 23 percent to 12.6 percent. Preterm and low birthweight levels both rose for 2002. The twin birth rate continued to climb, but the rate of triplet/+ births was down slightly.

Births: final data for 2002.

Embryonic stem cells have huge potential in the field of tissue engineering and regenerative medicine as they hold the capacity to produce every type of cell and tissue in the body. In theory, the treatment of human disease could be revolutionized by the ability to generate any cell, tissue, or even organ, 'on demand' in the laboratory. This work reviews the history of murine and human ES cell lines, including practical and ethical aspects of ES cell isolation from pre-implantation embryos, maintenance of undifferentiated ES cell lines in the cell culture environment, and differentiation of ES cells in vitro and in vivo into mature somatic cell types. Finally, we discuss advances towards the clinical application of ES cell technology, and some of the obstacles which must be overcome before large scale clinical trials can be considered.

https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1365-2184.2004.00298.x

Embryonic stem cells.

The effectiveness of blastocyst culture and transfer in human in-vitro fertilization (IVF) was evaluated in a prospective randomized trial in patients having a moderate to good response to gonadotrophin stimulation. Embryos were transferred either on day 3 after culture to around the 8-cell stage in Ham's F-10 medium supplemented with fetal cord serum, or on day 5 after culture to the blastocyst stage in the sequential serum-free media G 1.2 and G 2.2. The pregnancy rates after transfer on day 3 or day 5 were equivalent, 66 and 71% respectively; however, significantly more embryos were transferred on day 3 (3.7) than on day 5 (2.2). The number of blastocysts transferred did not affect the implantation rate, and pregnancy rates when either two or three blastocysts were transferred were 68 and 87% respectively. The implantation rate of the blastocysts (50.5% fetal heart beat) was significantly higher compared to the cleavage stage embryos transferred on day 3 (30.1%). The percentage of blastocyst development was not affected by the number of 2-pronuclear embryos, or by maternal age. Irrespective of the number of blastocysts formed, pregnancy rates were similar. Furthermore, the pregnancy rate following blastocyst transfer in patients with 10 or more follicles at the time of human chorionic gonadotrophin administration was not affected by patient age. More than 60% of patients having blastocyst culture and transfer had supernumerary embryos for cryopreservation. The establishment of a pregnancy following thaw and transfer confirmed the viability of cryopreserved blastocysts cultured in the absence of serum or co-culture. The ability to transfer just two blastocysts while maintaining high pregnancy rates will therefore help to eliminate high order multiple gestations and improve the overall efficiency of human IVF.

/pdf/a-prospective-randomized-trial-of-blastocyst-culture-and-j2g9jptfdm.pdf

A prospective randomized trial of blastocyst culture and transfer in in-vitro fertilization.

Objectives—This report presents 2004 data on U.S. births according to a wide variety of characteristics. Data are presented for maternal demographic characteristics including age, live-birth order, race, Hispanic origin, marital status, and educational attainment; maternal lifestyle and health characteristics (medical risk factors, weight gain, and tobacco use); medical care utilization by pregnant women (prenatal care, obstetric procedures, characteristics of labor and/or delivery, attendant at birth, and method of delivery); and infant characteristics (period of gestation, birthweight, Apgar score, congenital anomalies, and multiple births). Also presented are birth and fertility rates by age, live-birth order, race, Hispanic origin, and marital status. Selected data by mother's state of residence are shown, as well as data on month and day of birth, sex ratio, and age of father. Trends in fertility patterns and maternal and infant characteristics are described and interpreted. Methods—Descriptive tabulations of data reported on the birth certificates of the 4.1 million births that occurred in 2004 are presented.

/pdf/births-final-data-for-2004-2h3is32myf.pdf

Births: final data for 2004.

Culture and transfer of human blastocysts increases implantation rates and reduces the need for multiple embryo transfers

The pregnancy rate and implantation rate following blastocyst transfer in the human have been reported to be high; however, it has remained necessary to transfer 2-3 blastocysts to achieve these rates. Morphological criteria are currently used to select blastocysts for transfer and have some limited correlation with ongoing viability. Glucose metabolism of 189 human morula to blastocyst stage embryos was analysed using a non-invasive ultramicrofluorescence technique to determine if this could be used to predict viability. There was a linear trend to increased glucose uptake with progression from the morula to the hatching/hatched blastocyst stage of development, whereas glycolytic activity did not vary. There was no consistent difference in glucose uptake or glycolytic activity for embryos at the various morphological stages on day 5 compared to day 6 in vitro. Glucose uptake and glycolytic activity of the nine embryos positively identified as having implanted following transfer varied and were apparently not different from the values for embryos that failed to implant. In addition, viability was demonstrated to be compatible with high glycolytic activity, with four of nine implanted embryos having a glycolytic activity in the highest 15% of the population of embryos studied. Glucose uptake and glycolytic activity of male and female embryos did not appear to be different. Glucose metabolism cannot be used prospectively to select viable human morula or blastocyst stage embryos for transfer and it is also unlikely to be a useful tool to predict the sex of the embryo.

https://www.rbmojournal.com/article/S1472-6483(10)61980-3/pdf

Pauline Vella

Papers

Culture and transfer of human blastocysts increases implantation rates and reduces the need for multiple embryo transfers

Glucose metabolism of human morula and blastocyst-stage embryos and its relationship to viability after transfer.

Induction of glycolysis in the day-3 mouse embryo by glucose

Development of viable human embryos to the blastocyst stage following initial culture in a simple or a complex culture medium