The main aim of this study was to investigate the biological activities and immune modulation changes of chorionic villi mesenchymal stem cells (CV-MSC) after long term culture. The morphology of the CV-MSC of passage 3 and passage 9 were observed by microscopy, and their phenotypes were detected by flow cytometry. CV-MSC of passage 3 and 9 were co-cultured with PHA-stimulated PBMNC, and IFN-γ concentration in culture medium was detected by ELISA. The mRNA expression of COX-2, HGF and HLA-G in CV-MSC were detected by real-time PCR. The results showed that after long term culture, the CV-MSC kept the MSC morphology and most of the phenotypes including CD31, CD34, CD44, CD45, CD62L, CD73, CD90, CD105, CD117, CD151, CD235a, CD271 and HLA-DR, while the CD49d was significantly up-regulated. Immune modulation ability of CV-MSC was reduced and the mRNA expression of COX-2 and HGF was down regulated after long term culture, but the expression of HLA-G did not found to be obvious change. It is concluded that the long term in vitro expansion changes the expression of CD49d and reduces immune modulation of CV-MSC.
Cells, Cultured ; Chorionic Villi ; immunology ; Female ; Humans ; Integrin alpha4 ; metabolism ; Mesenchymal Stromal Cells ; cytology ; immunology ; Monocytes ; cytology ; Placenta ; cytology ; Pregnancy

OBJECTIVE: To determine the difference in aspartyl-(asparaginyl) beta-hydroxylase (AAH) expression level in villi between patients with missed abortion and normal women with early pregnancy, and to confirm the expression loci of AAH in villi. METHODS: A total of 50 patients of missed abortion were collected and categorized into a test group, which was subdivided into Group 1 and Group 2. Patients in Group 1 (n=20) were of confirmed etiological disorders while those in Group 2 (n=30) showed no obviously etiological clues. In addition, 20 women of early pregnancy with artificial abortion were categorized into a control group, whose embryos were sonographically confirmed alive before surgery. The 50 patients of missed abortion were also subdivided into a group within 4 weeks and a group over 4 weeks according to the time that the embryo stayed in utrine after death. Immunohistochemical technique and computer image analysis were used to detect the expression loci and the level of AAH in villi. RESULTS: AAH was expressed in the endochylema and nucleus of trephocyte both in missed abortion and normal early pregnancy. The expression level of AAH in villi of missed abortion was much lower than that of in villi of normal early pregnancy (P<0.05). The expression level had no difference between different groups of patients with missed abortion(P>0.05). CONCLUSION Low expression of AAH in the endochylema and nucleus of trephocyte may play a role in patients with missed abortion.
Abortion, Missed ; enzymology ; Adult ; Case-Control Studies ; Chorionic Villi ; enzymology ; Female ; Humans ; Mixed Function Oxygenases ; genetics ; metabolism ; Pregnancy

OBJECTIVETo explore the differences of metabolic footprint in the conditioned culture medium of placental explants between early-onset and late-onset severe preeclampsia.

METHODSIn 13 cases of early-onset severe preeclampsia and 14 cases of late-onset severe preeclampsia, the placentas were sampled at the surface of the maternal placenta. High performance liquid chromatography-mass spectrometry (HPLC-MS) was used to determine the differences in the metabolites in the conditioned culture medium of the placental villous explants cultured in 6% atmospheric O(2) for 96 h. Standard samples were used to establish the tryptophan and kynurenine chromatography library by HPLC-MS to analyze the concentration of tryptophan and kynurenine in the conditioned culture medium.

RESULTSThirty-six metabolites showed statistically significant differences between early-onset and late-onset severe preeclampsia (P<0.05). The concentration of kynurenine was significantly higher in early-onset severe preeclampsia than in late-onset severe preeclampsia (P<0.05).

CONCLUSIONEarly-onset and late-onset severe preeclampsia may have different pathogeneses. By detecting the concentration of metabolites, metabolomic strategies provide a new means for predicting the onset time of severe preeclampsia.

Chorionic Villi ; metabolism ; Culture Media, Conditioned ; chemistry ; Female ; Humans ; In Vitro Techniques ; Kynurenine ; metabolism ; Ornithine ; metabolism ; Placenta ; metabolism ; Pre-Eclampsia ; metabolism ; Pregnancy ; Tryptophan ; metabolism

Chorionic Villi ; metabolism ; pathology ; ultrastructure ; Diabetes, Gestational ; metabolism ; pathology ; Female ; Glucose ; metabolism ; Glycogen ; metabolism ; Humans ; Lipids ; analysis ; Microscopy, Electron, Scanning ; Microscopy, Electron, Transmission ; Placenta ; metabolism ; pathology ; ultrastructure ; Pregnancy

OBJECTIVETo explore the effects of 50 Hz sinusoidal magnetic fields (MF) on secretion function of primary human villous trophoblasts in vitro, and the interference effect of "noise" MF.

METHODSThe trophoblasts were isolated from human villus by trypsin digestion and incubated in DMEM medium.Then the trophoblasts were exposed to 0.4 mT 50 Hz MF and/or "noise" MF respectively for different durations. Each exposure group was matched with one control group which was from the same villus and cultured with the same condition except the MF exposure. The concentrations of human chorionic gonadotropin (HCG) and progesterone in the culture medium were measured by immunofluorescence. Statistical significance of differences between means was determined by one way-ANOVA with P<0.05 considered significant.

RESULT50 Hz MF inhibited the HCG and progesterone secretion significantly when exposure for 72 h (compared with control group, P<0.05). There was no significant change of HCG and progesterone secretion when trophoblasts were exposed to 0.4 mT "noise" MF within 72 h (compared with control group, P>0.05). However, by superimposing the "noise" MF, the inhibition of HCG and progesterone secretion of trophoblasts induced by 50 Hz MF was eliminated.

CONCLUSIONThe exposure to 50 Hz MF for long period could inhibit trophoblasts secreting HCG and progesterone, and the "noise" MF with the same intensity could eliminate the effects induced by 50 Hz MF.

Biological Transport ; radiation effects ; Bodily Secretions ; radiation effects ; Cells, Cultured ; Chorionic Gonadotropin ; metabolism ; Chorionic Villi ; metabolism ; radiation effects ; DNA ; radiation effects ; Electromagnetic Fields ; Humans ; Noise ; Progesterone ; metabolism ; Trophoblasts ; metabolism ; radiation effects

OBJECTIVETo investigate the expression of vascular endothelial growth factor (VEGF) in human placental trophoblasts and the role of VEGF in regulating placental villous angiogenesis.

METHODSPlacental samples were obtained from 10 pregnant women receiving induced abortion in the first trimester, 10 receiving induced labor in the second trimester and 10 having cesarean section at term delivery, with gestational duration of 6-9, 18-22 and 37-38 weeks, respectively. All the samples were fixed in formalin solution and prepared for the morphological study. The expression of VEGF and vascular distribution in the placental villi were examined and evaluated by immunohistochemistry and stereomorphometry, respectively.

RESULTSIn the course of pregnancy, there was a significant decrease in the level of VEGF expression in chorionic villi (28.19+/-3.01, 18.65+/-2.43, 4.95+/-0.86, respectively, P<0.01). The radial parameters of the blood vessels showed no significant changes (26.67+/-7.74, 25.08+/-4.67, 23.36+/-5.30, respectively, P>0.05), but the length density of the blood vessels increased significantly (1.46+/-0.64, 5.58+/-1.31, 19.56+/-1.40, respectively, P<0.01).

CONCLUSIONDuring gestation, VEGF expression in chorionic villi gradually weakens but the length density of the blood vessels increases, indicating that VEGF is not the only regulatory factor of angiogenesis in the chorionic villi.

Adult ; Chorionic Villi ; blood supply ; Female ; Gestational Age ; Humans ; Neovascularization, Physiologic ; physiology ; Placenta ; metabolism ; Pregnancy ; Trophoblasts ; metabolism ; Vascular Endothelial Growth Factor A ; biosynthesis

OBJECTIVETo investigate the relationship between fetal chromosomal karyotype and early spontaneous abortion, and the effect of the environmental factors on spontaneous abortion.

METHODSChoronic villi from 252 cases of missed abortion were sampled as patient group and 50 normal pregnancies as control group. Chorionic villi were cultured and karyotype analysis was performed by G-banding. Clinical information was collected.

RESULTSThe rate of chorion chromosome abnormality in the patient group was 58.09%, significantly higher than that in the control group (4.17%) (P<0.01). Among the 140 cases of karyotype abnormalities, 81 were trisomy, 29 were monosomy X and 17 were polyploidy, accounting for 57.86%, 20.71% and 12.14% of total abnormalities, respectively. Long time and low dose radiation exposure of the pregnant female seemed to be related with spontaneous abortion(P<0.01).

CONCLUSIONChorion chromosome abnormality is a major reason of early spontaneous abortion and karyotype analysis of chorionic villus is of clinical importance. For pregnant women, long-term exposure to computers and television seems a risk factor for missed abortion.

Abortion, Missed ; genetics ; Adolescent ; Adult ; Case-Control Studies ; Chorionic Villi ; metabolism ; Chromosome Aberrations ; Female ; Humans ; Karyotyping ; Middle Aged ; Pregnancy ; Risk Factors ; Young Adult

OBJECTIVE: To investigate the expression of Talin1 in the fallopian tube and chorionic villi in patients with tubal pregnancy and its role in regulating invasion and migration of trophoblasts. METHODS: Immunohistochemistry and Western blotting were used to detect the localization and expression level of Talin1 in the fallopian tube and chorionic villi in patients with tubal pregnancy and in women with normal pregnancy. In the cell experiment, HTR-8/SVneo cells was transfected with Talin1 siRNA and the changes in cell invasion and migration were assessed using scratch assay and Transwell assay. The expressions of MMP-2, MMP-9, N-cadherin and Snail in the transfected cells were detected by qRT-PCR and Western blotting. RESULTS: Positive expression of Talin1 was detected in both normal fallopian tube tissues and tissues from women tubal pregnancy, and its expression was localized mainly in the cytoplasm of cilia cells. The expression level of Talin1 was significantly higher in both the fallopian tube and chorionic villi in women with tubal pregnancy than in normal fallopian tube and chorionic villi samples (P < 0.01). In HTR-8/SVneo cells, transfection with Talin1 siRNA significantly inhibited cell invasion (P < 0.01) and migration (P < 0.05), down-regulated the expression of N-cadherin, MMP-2 and Snail (P < 0.05), and up-regulated the expression of MMP-9 in the cells (P < 0.05). CONCLUSION The expression of Talin1 in the fallopian tube and chorionic villi is significantly increased in women with tubal pregnancy, suggesting the association of Talin1-regulated trophoblast cell invasion with the occurrence of tubal pregnancy.
Cadherins/metabolism* ; Cell Movement ; Chorionic Villi/metabolism* ; Fallopian Tubes/metabolism* ; Female ; Humans ; Matrix Metalloproteinase 2/metabolism* ; Matrix Metalloproteinase 9/metabolism* ; Pregnancy ; Pregnancy, Tubal/metabolism* ; RNA, Small Interfering/metabolism* ; Talin/metabolism* ; Trophoblasts/metabolism*

OBJECTIVETo investigate the expression of TM4SF9 in the villi of early pregnancy, hydatidiform mole, invasive hydatidiform mole and chorionic carcinoma tissue.

METHODSImmunohistochemistry was used to detect the expression of TM4SF9 in normal villi of early pregnancy, hydatidiform mole, invasive hydatidiform mole and chorionic carcinoma tissues.

RESULTSTM4SF9 was expressed in the cytotroblasts but not in the syncytiotrophoblast of normal villi. The intensity of TM4SF9 expression increased in the order of normal villi, hydatidiform mole, invasive hydatidiform mole and chorionic carcinoma, with strong positivity rates of 0, 10%, 36.4% and 100%, respectively, showing significant differences between the samples (P<0.001).

CONCLUSIONTM4SF9 expression in the trophoblasts may relate to their invasiveness and play an important role in the metastasis of trophoblastic tumor.

Adult ; Choriocarcinoma ; metabolism ; Chorionic Villi ; metabolism ; Female ; Humans ; Hydatidiform Mole ; metabolism ; Hydatidiform Mole, Invasive ; metabolism ; Immunohistochemistry ; Membrane Proteins ; biosynthesis ; Pregnancy ; Tetraspanins ; Trophoblastic Neoplasms ; metabolism ; Trophoblasts ; metabolism ; Uterine Neoplasms ; metabolism

OBJECTIVEMucopolysaccharidosis type II (MPS II, Hunter syndrome, OMIM 309900) is an X-linked recessive lysosomal storage disease resulting from a deficiency of iduronte-2-sulphate sulphatase (IDS). The present study aimed to establish an enzyme assay method for IDS activity for carrying out postnatal and prenatal diagnosis of MPS II by means of IDS activity assay on plasma, uncultured chorionic villi (CV) and cultured amniotic fluid cells (AF cell) using a new synthesized substrate.

METHODSA fluorigenic substrate (4-methylumbelliferyl-alpha-iduronate-2-sulphate, MU-alpha-Idu-2S) was used for the assay of IDS activity. IDS activity in plasma was determined for diagnosis of the proband. Prenatal diagnosis in 10 pregnancies at risk was carried out according to IDS activity on uncultured CV at 11th week or on cultured AF cell at 18th week of gestation. At the same time, IDS activity was also determined in the maternal plasmas to observe the change of IDS activity in pregnancy. The fetal sex determination was performed by PCR amplification of the ZFX/ZFY genes.

RESULTThe IDS activity in plasma of normal controls and obligate heterozygotes were 240.2 - 668.2 nmol/(4 hxml) and 88.7 - 547.9 nmol/(4 hxml), respectively, while the enzyme activities in plasmas were in the range of 0.3 - 18.6 nmol/(4 hxml) in affected male. The IDS activities were 37.2 - 54.9 nmol/(4 hxmg protein) and 21.4 - 74.4 nmol/(4 hxmg protein) in CV and cultured AF cells respectively. Out of 50 suspected cases, 46 were diagnosed as having MPS II and 4 were excluded. Prenatal diagnosis was performed on 10 pregnancies at risk. Four of 5 male fetuses [IDS activity were 4.7, 1.8, 7.0 nmol/(4hxmg protein) in CV, 0.6 nmol/(4 hxmg protein) in AF cell] were diagnosed as having MPS II and the other 5 fetuses were normal females [IDS activity were: 48.7, 5.9, 25.2 nmol/(4 hxmg protein) in CV, 55.2, 40.9 nmol/(4 hxmg protein) in AF cell]. Increased IDS activity was observed in plasma of the pregnant women with unaffected fetuses, while the IDS activity decreased in pregnancies with affected fetuses. IDS activity of one female fetus was very low [5.9 nmol/(4 hxmg protein)], but the IDS activity in maternal plasmas increased, this fetus was a normal female.

CONCLUSIONSThe method using a synthesized fluorigenic 4-methylumbelliferyl-substrate was a sensitive, rapid and convenient assay of IDS activity and was reliable for early prenatal diagnosis. Determination of fetal sex would be helpful in excluding the female fetus with low IDS activity from being considered as an affected male fetus. It would be further helpful if IDS activity in maternal plasma was taken into account.

Amniotic Fluid ; cytology ; enzymology ; Cells, Cultured ; Child ; Child, Preschool ; China ; epidemiology ; Chorionic Villi ; enzymology ; Chorionic Villi Sampling ; Enzyme Assays ; methods ; Female ; Fetus ; enzymology ; Fluorometry ; methods ; Heterozygote ; Humans ; Hymecromone ; analogs & derivatives ; Iduronate Sulfatase ; blood ; metabolism ; Iduronic Acid ; analogs & derivatives ; Karyotyping ; Male ; Mucopolysaccharidosis II ; diagnosis ; enzymology ; epidemiology ; Polymerase Chain Reaction ; Pregnancy ; Pregnancy, High-Risk ; blood ; Prenatal Diagnosis ; methods ; Reference Values ; Sex Factors