Duration of storage of cryopreserved human embryos.

The incidence of cell injury, embryo survival, and implantation following cryopreservation of zygotes and two- to five-cell embryos was studied in 100 patients in order to evaluate the effect of duration of storage. The incidence of individual cell survival was 58% regardless of the length of time kept in liquid nitrogen or the stage of the embryo at freezing. There were 104 of 208 (50%) thawed embryos that survived completely intact, and of those, 24 implanted successfully.

Cryopreservation of zygotes and early cleaved human embryos.

Zygotes and 2- to 5-cell human embryos were frozen in 1,2-propanediol and sucrose; results of the first 50 cycles (45 patients) are presented. A total of 41 zygotes (17 attempts at thawing) were thawed, resulting in six singleton clinical pregnancies (15% per embryo; 35% per cycle), of which three delivered, one aborted, and two are ongoing. Fifty-seven cleaved embryos were thawed in 33 other cycles, resulting in four singleton and one twin pregnancy (11% per embryo; 15% per cycle), of which four delivered and one is ongoing.

Are ultrasonic-guided follicular aspiration and flushing safe for the oocyte?

Oocyte collection was carried out in 53 patients by ultrasonically guided abdominal puncture under local or epidural anesthesia. Follicles were aspirated and flushed two to six times using a syringe with culture medium. A total of 196 oocytes was collected and 84 of those (42.

Transfer of immunity to Ostertagia circumcincta and IgA memory between identical sheep by lymphocytes collected from gastric lymph.

Sheep rendered immune to Ostertagia circumcincta were challenged with 50,000 larvae and lymphocytes were collected from the gastric lymph up to eight days after challenge. The cells were transferred intravenously to genetically identical worm-free sheep which, together with controls, were challenged with 50,000 larvae and killed nine days later. Cells obtained during the donors lymphoblast response to challenge transferred partial immunity, measured either as stunting or loss of worms.

Factors affecting survival and implantation of cryopreserved human embryos.

Five- to ten-cell embryos and expanded blastocysts from 68 patients were thawed in an attempt to establish pregnancy. Three fresh embryos had been replaced unsuccessfully in these patients. Forty-five patients had intact freeze-thawed embryos replaced and 12 became clinically pregnant.

Cryopreservation of cleaving embryos and expanded blastocysts in the human: a comparative study.

The survival and implantation capacity of cryopreserved cleaving (5-cell to 10-cell) human embryos and expanded blastocysts was compared. Twice as many cleaving embryos were frozen as were expanding blastocysts because of the low developmental potential of human embryos in vitro. However, significantly more expanded blastocysts survived cryopreservation than cleaving embryos, and relatively more pregnancies were established by the replacement of thawed blastocysts than by the replacement of thawed cleaving embryos.

Treatment of idiopathic infertility, cervical mucus hostility, and male infertility: artificial insemination with husband's semen or in vitro fertilization?

Couples with male infertility (n = 86), idiopathic infertility (n = 68), and cervical mucus hostility (n = 48) of a long duration were treated either by in vitro fertilization (IVF) or artificial insemination with husband's semen (AIH). The incidence of pregnancy per cycle in couples in whom the male partner was infertile was significantly (P less than 0.01) higher after IVF, compared with AIH (21% versus 5%, respectively).

Implantation, abortion, and birth after in vitro fertilization using the natural menstrual cycle or follicular stimulation with clomiphene citrate and human menopausal gonadotropin.

The incidence of pregnancy and abortion was analyzed in 1679 patients having embryos replaced after oocyte recovery and in vitro fertilization in order to alleviate their infertility. In these patients, 364 pregnancies were achieved and 108 abortions occurred. Patients were treated either on their natural cycle, having a spontaneous luteinizing hormone (LH) surge to induce ovulation, or after ovarian stimulation using clomiphene citrate alone or in combination with human menopausal gonadotropin (hMG).

Prolonged storage of human spermatozoa at room temperature or in a refrigerator.

Spermatozoa from patients who underwent in vitro fertilization (IVF) therapy were prepared free from seminal plasma with the use of IVF culture medium supplemented with 8% human serum. Samples were then stored either at room temperature or in a refrigerator, and their motility and ability to penetrate zona-free hamster eggs were assessed every second day. With storage at room temperature, motility declined by less than half over the first 14 days, with some samples still active 20 days after preparation.

Pregnancies following the frozen storage of expanding human blastocysts.

Human blastocysts were frozen in Earle's solution containing pyruvate and human serum, using glycerol as cryoprotectant, and stored in liquid nitrogen. Thawed blastocysts were replaced in 11 patients, which resulted in two pregnancies. One blastocyst giving a pregnancy was hatching when replaced.

Cytogenetic and blood group studies of sheep/goat chimaeras.

Aggregation chimaeras were composed of quarter (or 1 cell) contributions from 4-cell blastocysts of sheep or goats, or of an 8-cell blastocyst of one species enveloped in three 8-cell blastocysts of the other. Gestation was in sheep or goat recipient females. Of the 10 living animals born, 3 were identified as interspecific chimaeras by body conformation and coat type among the 7 quarter/quarter aggregations and 1 among the 3 giant aggregates.

In vitro fertilization using cryopreserved donor semen in cases where both partners are infertile.

The incidence of pregnancy after in vitro fertilization (IVF) was studied in a group of 38 couples (55 cycles) where both partners were infertile. Cryopreserved donor semen (IVF-D) was used in all cycles. Results were compared with those in a control group of couples where the husband's semen was considered normal and only the wife was infertile.

In vitro fertilization: a treatment for male infertility.

Three-quarters of 41 women whose husbands were the cause of infertility in the couple had one or more embryos replaced after in vitro fertilization. One-third became pregnant despite their long history of infertility; the incidence of pregnancy was 45% after one or more replacements of embryos. In a group of 61 couples where both partners were infertile, 26% became pregnant.

Treatment of male infertility by in vitro fertilization: factors affecting fertilization and pregnancy.

IVF was attempted in more than 100 couples where the man was considered to be the cause of the infertility. 57% of all oocytes were fertilized and embryo replacement was achieved in 60-70% of cases. Pregnancies following IVF were established in cases with a long duration of infertility and different male factors such as oligospermia, teratospermia, asthenospermia and auto-immunity.

Resistance to Haemonchus contortus transferred between genetically histocompatible sheep by immune lymphocytes.

The pattern of faecal egg counts after infection with 10,000 Haemonchus contortus larvae was similar within but not between, four pairs of monozygous twin sheep. Transfer of whole lymph or washed lymph cells from three immune donor sheep to their identical co-twin recipients reduced the susceptibility of the recipients to challenge with 10,000 larvae as measured by faecal egg counts. Cells from a nonimmune donor did not have this effect.

The application of the zona-free hamster egg test for the prognosis of human in vitro fertilization.

The zona-free hamster egg test was carried out using spermatozoa from 15 men which consistently failed to fertilize their wives' oocytes in vitro. Spermatozoa from nine of these men fertilized hamster eggs in vitro, indicating that positive results in this assay are an unreliable guide to human in vitro fertilization. Donor spermatozoa were needed to fertilize the wife's oocytes in three of these cases.

Factors influencing the success of in vitro fertilization for alleviating human infertility.

The program for in vitro fertilization at Bourn Hall began in October 1980. Various types of infertility have been treated during this time using the natural menstrual cycle or stimulation of follicular growth with antiestrogens and gonadotrophins. Follicular growth and maturation are assayed by urinary estrogens and LH, monitored regularly during the later follicular stage.

Interspecific chimaerism between sheep and goat.

In rodents, chimaeric blastocysts produced by combining embryonic cells of two different species have been used in investigations of cell lineage and interaction during development (Mus musculus-Rattus norvegicus, M. musculus-Clethrionomys glareolus, M. musculus-Mus caroli).