1.Expression of macrophage colony-stimulating factor in human luteinized granulosa cells
Zhifen ZHANG ; Salmassi ALI ; Mettler LISELOTTE
Chinese Journal of Obstetrics and Gynecology 2001;0(08):-
Objective To detect and localize the expression of macrophage colony stimulating factor (M CSF) in human luteinized granulosa cells and investigate the effects of follicle stimulating hormone (FSH) on M CSF production by human luteinized granulosa cells in vitro Methods Human luteinized granulosa cells were isolated from follicular fluid of superovulated infertile patients undergoing intracytoplasmatic sperm injection Some of the luteinized granulosa cells were used for detecting M CSF by immunocytochemical staining (ABC method) Most of them were cultured with HAM′s F 10 medium alone or plus various concentrations of FSH (2, 5, 15, 25 IU/ml) The media were collected after 72 hours and their M CSF contents were measuered by a solid phase enzyme linked immunosorbant assay Results About 80% of the luteinized granulosa cells were positively stained for M CSF antibody The immunoreactive signals of M CSF were specially located in the cytoplasma of the luteinized granulosa cells No immunoreactivity of M CSF was observed in vaginal squamous epithelial cells through contaminations during transvaginal oocyte retrieval The concentration of M CSF in cultured luteinized granulosa cells medium without FSH stimulation was low However, addition of 2, 5, 15, 25 IU/ml concentrations of FSH caused significant dose dependent increases of M CSF production after 72 hours culture by luteinized granulosa cells ( P
2.Study on the mechanism of oocyte aneuploidy by spectral karyotyping.
Wen CHEN ; Gui-jin ZHU ; Liselotte METTLER
Chinese Journal of Medical Genetics 2005;22(4):411-414
OBJECTIVETo establish the method of detecting oocyte aneuploidy by spectral karyotyping (SKY).
METHODSThe unfertilized oocytes were fixed 1-2 days after oocyte retrieval. Spectral karyotyping was performed according to the protocol.
RESULTS64% of oocytes were normal, 36% of oocytes were aneuploidy, of which 22% were due to nondisjunction and 14% unbalanced predivision.
CONCLUSIONSKY is an effective method for detecting oocyte aneuploidy. Both nondisjunction and unbalanced predivision are involved in oocyte aneuploidy formation.
Aneuploidy ; Female ; Humans ; Male ; Oocytes ; cytology ; metabolism ; Pregnancy ; Reproducibility of Results ; Spectral Karyotyping ; methods
3.Study on the mechanism of oocyte aneuploidy formation by multicolor fluorescence in situ hybridization.
Chinese Journal of Medical Genetics 2004;21(3):203-206
OBJECTIVETo investigate the mechanism of oocyte aneuploidy formation.
METHODSThe unfertilized oocytes were fixed 1-3 days after oocyte retrieval. Multicolor fluorescence in situ hybridization (M-FISH) was performed according to the Vysis protocol to check the chromosome status in oocytes by using centromeric enumerator probes for chromosomes 16, 18 and locus-specific probes for chromosomes 13, 21 and 22.
RESULTS47% oocytes were found to be normal, while 53 % oocytes were abnormal. Nondisjunction was found in 22(18%) oocytes, unbalanced predivision of chromatids in 15(12%) oocytes, balanced predivision of chromatids in 45(36%) oocytes. The balanced predivision rate in oocytes aged in vitro>24h was much higher than that in oocytes aged in vitro CONCLUSIONBoth nondisjunction, balanced and unbalanced predivision of chromatids are involved in the oocyte aneuploidy formation. Balanced predivision is related to the time in culture.
Adult
;
Aneuploidy
;
Female
;
Humans
;
In Situ Hybridization, Fluorescence
;
methods
;
Karyotyping
;
Oocytes
;
ultrastructure