Improvements in assessment of human failed fertilization oocytes using the supravital dye Hoechst 33342.
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Benzimidazoles↗
Biomedical subjects
J Witmyer
Publications and source records attributed to J Witmyer.
Effects of human serum and plasma on development of mouse embryos in culture media.
Mouse embryos were cultured in Ham's F-10 medium (unsupplemented) and Ham's F-10 medium supplemented with either 15% fetal cord serum, 15% fetal cord plasma, 15% maternal serum, or 15% maternal plasma. None of the blood supplements significantly increased the numbers of embryos that developed to blastocysts, and the last three of the blood supplements inhibited embryo development. The cause of the inhibition was not identified. Estradiol concentrations of blood samples used as media supplements were found not to correlate with inhibition of embryo.
Animals↗
Evaluation of human fetal cord sera, Ham's F-10 medium, and in vitro culture materials with a mouse in vivo fertilization system.
A total of 54 human fetal cord sera, 42 lots of Ham's F-10 medium, and 36 lots of plastics were tested during a 10-month period with a mouse in vivo fertilization system. Two-cell embryos were collected from the oviducts of superovulated and mated (C57BL/6 X CBA)F1 mice. After collection, 2-cell embryos were distributed among test media and a modified Krebs-Ringer control medium. The test material passed quality control standards if a minimum of 80% of the original 2-cell embryos reached the blastocyst stage. Of the 54 cord sera examined, 8 (15%) were below our standards; 6 (14%) of the Ham's F-10 media and 4 (11%) of the plastics failed to pass minimum requirements. In all cases, the failed materials promoted slower-growing mouse embryos and increased the number of degenerates. It is our opinion that the mouse in vivo fertilization system has a valuable place in any in vitro fertilization program.
Animals↗
Maturation and fertilization of morphologically immature human oocytes in a program of in vitro fertilization.
Oocytes of varying stages of maturity were aspirated from follicles primed with either human menopausal gonadotropin (hMG) and human chorionic gonadotropin (hCG) or a combination of follicle-stimulating hormone (FSH), hMG and hCG. Of the aspirated oocytes from 44 cycles, 74 were considered to be immature by virtue of morphologic characteristics of the oocytes and the degree of intercellular expansion of the associated cumular and membrana granulosa cells. After incubation periods of 22 to 35 hours in a Ham's F-10-based culture medium, these immature oocytes were inseminated with sperm donated by the patient's husband. Ultimately, 44 conceptuses were transferred to the respective uteri of 30 patients. Eight pregnancies were established as a result of these 30 transfers, two of which resulted from the transfer of only developed immature oocytes.
Chorionic Gonadotropin↗
Vital initiation of pregnancy (VIP) using human menopausal gonadotropin and human chorionic gonadotropin ovulation induction: Phase I--1981.
Laparoscopies for oocyte aspiration in 31 cycles were performed on 25 patients receiving human menopausal gonadotropin and human chorionic gonadotropin. Sixty oocytes were aspirated, of which 48 were considered preovulatory. Ninety-seven percent (58 of 60) of the oocytes were found in the original aspirate, and the remaining oocytes were found in either the first or second follicle wash. The fertilization rate per preovulatory oocyte was 33% (16 of 48), whereas on a per cycle basis it was 39% (12 of 31). A total of 15 conceptuses (2-cell = 5; 3-cell = 3; 4-cell = 7) were transferred to 12 patients, and two pregnancies were established. These pregnancies were established by transfers of 3-cell and 4-cell conceptuses at approximately 47 hours after insemination. Both pregnancies resulted in term deliveries of normal infants.
Cell Separation↗
Vital initiation of pregnancy (VIP) using human menopausal gonadotropin and human chorionic gonadotropin ovulation induction: phase II--1981.
In a program for in vitro fertilization, laparoscopies for oocyte aspiration were performed on 24 patients receiving human menopausal gonadotropin and human chorionic gonadotropin. Of the 40 preovulatory oocytes that were recovered from these patients, 33 (83%) were fertilized and 30 (75%) cleaved and were transferred. Ten immature oocytes were collected, and attempts were made to mature these in vitro prior to insemination. All ten oocytes (100%) did fertilize, and seven (70%) cleaved and were transferred. Morphologic variation was noted between cleaving conceptuses, even in those conceptuses responsible for establishing pregnancies. Five pregnancies resulted from 19 embryo transfers (26%).
Chorionic Gonadotropin↗
The program for in vitro fertilization at Norfolk.
Several aspects of the program of in vitro fertilization (IVF), or, as it is called in Norfolk, the program for the Vital Initiation of Pregnancy (VIP), have been or are in the process of publication. However, because there has been no overall account, it seems appropriate to give a brief report of a general nature covering the period from the beginning of the effort in late February 1980 through December 31, 1981. Although minor changes were constantly made in the protocol, there were two major revisions. Therefore, a discussion of the program during three distinct periods, i.e., 1980, 1981-Phase I, and 1981-Phase II, is necessary. During 1980 and 1981 all patients had either no fallopian tubes or irreparable tubes.
Adolescent↗