Jiaen Liu

High fertilization rate obtained after intracytoplasmic sperm injection with 100% nonmotile spermatozoa selected by using a simple modified hypo-osmotic swelling test

OBJECTIVE To report a high fertilization rate after intracytoplasmic sperm injection (ICSI) in patients with 100% nonmotile spermatozoa selected by a simple modified hypo-osmotic swelling test. DESIGN Clinical study. SETTING Hospital-based IVF center. PATIENT(S) Three couples with infertility due to asthenospermia. INTERVENTION(S) The hypo-osmotic swelling test with 150-mOsm NaCl solution was used to select viable spermatozoa before ICSI. Three patients provided semen samples and one of these three also had a testicular biopsy. MAIN OUTCOME MEASURE(S) Selection of viable spermatozoa using the hypo-osmotic swelling test with 150-mOsm NaCl solution for ICSI. RESULT(S) No motile spermatozoa were found in three ejaculated semen samples and one testicular biopsy. Fifty-seven metaphase-II oocytes were injected with hypo-osmotic swelling test-positive ejaculated or testicular spermatozoa. Fifty-five (96.5%) of these oocytes were intact after injection. Forty-two (76.4%) of 55 oocytes showed two pronuclei, and 40 of the 42 fertilized oocytes cleaved. One patient had all embryos cryopreserved because of the risk of hyperstimulation; two other patients had embryos transferred. One ongoing pregnancy resulted. CONCLUSION This hypo-osmotic swelling test with 150-mOsm NaCl solution is a simple and efficient method for selection of viable spermatozoa. A high fertilization rate can be obtained using ICSI with viable spermatozoa selected by using this hypo-osmotic swelling test.

Successful in vitro maturation of human oocytes not exposed to human chorionic gonadotropin during ovulation induction, resulting in pregnancy

OBJECTIVE To report a case of successful in vitro maturation of human oocytes not exposed to hCG during ovulation induction, resulting in pregnancy after transfer of a frozen-thawed embryo resulting from intracytoplasmic sperm injection (ICSI) of the in vitro-matured human oocytes. DESIGN Clinical study. SETTING Hospital-based private IVF center. PATIENT(S) A couple with infertility due to severe oligospermia. INTERVENTION(S) Five immature oocytes were retrieved from a patient who failed to use hCG during ovarian stimulation and were cultured for 48 hours in B2 medium containing FSH and hCG. Five oocytes extruded the first polar body and underwent ICSI with frozen-thawed husband spermatozoa. MAIN OUTCOME MEASURE(S) In vitro maturation, fertilization after ICSI, embryo development, and pregnancy. RESULT(S) All five oocytes extruded the first polar body and were injected using ICSI. Three oocytes were fertilized, but two showed three pronuclei. The remainder, a two-pronuclei embryo subsequently cleaved and was cryopreserved. An ongoing pregnancy was obtained after-the transfer of this frozen-thawed embryo. CONCLUSION(S) Immature human oocytes at the germinal-vesicle stage that have not been exposed to hCG during ovarian stimulation can be matured in vitro and a normal pregnancy can result from ICSI of the in vitro-matured oocytes.

Effect of human hydrosalpinx fluid on murine embryo development and implantation

OBJECTIVE To determine the effect of hydrosalpinx fluid-containing medium on murine embryo development and implantation. DESIGN The development of one-, two-, and four-cell mouse embryos in medium containing 5%, 10%, and 20% of human hydrosalpinx fluid was observed. Implantation rates of mouse embryos transferred into the uterine horn with hydrosalpinx fluid-containing media were determined. SETTING Private hospital-based fertility center and IVF program. MAIN OUTCOME MEASURE(S) Percentage of embryos continuing cell division and implantation rates after ET. RESULT(S) Hydrosalpinx fluid in culture medium affected embryo development in a dose-dependent fashion. The injection of hydrosalpinx fluid-containing medium into the uterine horn did not affect embryo implantation. CONCLUSION(S) Hydrosalpinx fluid negatively affects murine embryo development, but its presence in the uterine horn at ET did not affect implantation.

Potential use of repeated fluorescence in situ hybridization in the same human blastomeres for preimplantation genetic diagnosis

OBJECTIVE To assess the feasibility of repeated fluorescence in situ hybridization (FISH) procedures in the same nucleus of a human blastomere. DESIGN Three consecutive FISH procedures were performed in the same human blastomere by using direct label fluorescence CEP and WCP probes (Vysis). SETTING Hospital-based private IVF program. PATIENT(S) Twenty-eight infertile couples who underwent conventional IVF in our center. INTERVENTION(S) Embryos from oocytes with three pronuclei after in vitro insemination were used in this study. MAIN OUTCOME MEASURE(S) The rates of nuclear loss, present signals, and absent signal were examined. RESULT(S) In group 1, the rates of presence of signals were 94% after the first FISH, 92% after the second FISH, and 88% after the third FISH. In group 2, the rates of presence of signals were 96% after the first FISH, 93% after the second FISH, and 87% after the third FISH. There was no statistically significant difference in the rates of nuclear loss, present signals, and absent signal between three consecutive FISH procedures and between CEP and WCP probes. CONCLUSION(S) Six or more chromosomes of a single blastomere may be examined with use of this repeated FISH procedure, which may be important for preimplantation genetic diagnosis.

Ultrarapid detection of sex chromosomes with the use of fluorescence in situ hybridization with direct label dna probes in single human blastomeres, spermatozoa, amniocytes, and lymphocytes

OBJECTIVE To assess the ultrarapid fluorescence in situ hybridization (FISH) procedure with a 1-minute hybridization time for gender determination. DESIGN Fluorescence in situ hybridization with direct label fluorescence DNA probes for chromosomes X and Y were tested with the use of different hybridization times and different cell types. SETTING Hospital-based IVF program. INTERVENTION(S) The efficiency of the FISH procedure with different hybridization times was compared with the use of male lymphocytes. The same FISH procedure, but with only 1-minute hybridization, was carried out in human blastomeres, spermatozoa, uncultured amniocytes, male lymphocytes, and female lymphocytes. MAIN OUTCOME MEASURE(S) Percentages of nuclei with positive signals. RESULT(S) The percentages of nuclei with positive signals in lymphocytes with hybridization times of 1, 3, 4, 10, 30, and 45 minutes were 97%, 97%, 98%, 98%, 98%, and 98%, respectively. The percentages of nuclei with positive signals after FISH with a 1-minute hybridization time in single blastomeres, spermatozoa, amniocytes, male lymphocytes, and female lymphocytes were 94%, 96%, 96%, 98%, and 97%, respectively. CONCLUSION(S) Chromosomes X and Y of human blastomeres. spermatozoa, uncultured amniocytes, and lymphocytes can be detected rapidly with the use of this ultrarapid FISH procedure with a 1-minute hybridization time.