人胎盘滋养细胞分选技术研究进展

人胎盘滋养细胞(TB)包括绒毛细胞滋养细胞(VCT)、合体滋养细胞(VST)和绒毛外细胞滋养细胞(EVT)。各种TB形态差异较大,在母-胎界面的功能也各不相同,此外由于母体细胞的污染,因此有必要对TB的类别加以区分。如何在保证细胞活性的同时提高纯度成为构建TB细胞系的主要困难之一。梯度离心、磁珠分选和流式细胞术是广泛应用的细胞分选技术。各种分选技术在TB研究中各有利弊,磁珠分选结合流式细胞术是目前较为有效的分选策略。     

激活素A对早孕胎盘细胞滋养细胞凋亡的作用

激活素A是转换生长因子13家族中的一员。研究表明，激活素A在细胞增殖、凋亡以及癌变过程中都有重要的调节作用；其中激活素A对具有增殖功能的细胞可诱导其发生凋亡。人的细胞滋养细胞是具有增殖能力的正常细胞，细胞膜上有激活素A受体表达，在妊娠期与激活素A结合，继而影响细胞滋养细胞增殖与凋亡。本研究旨在探讨激活素A对细胞滋养细胞凋亡的调节作用，报道如下。     

5-氟尿嘧啶联合顺铂、长春新碱治疗妊娠滋养细胞肿瘤的疗效分析

目的:探讨5-氟尿嘧啶(5-FU)微量泵持续泵入联合长春新碱、顺铂(FPV方案)治疗妊娠滋养细胞肿瘤(GTN)的临床疗效。方法:回顾分析2013年10月至2018年10月于郑州大学第二附属医院接受FPV(FPV组)及FAV(FAV组)化疗方案的82例GTN患者的临床资料及治疗结局。结果:82例患者中位随访时间37个月(8~70个月)。FPV组40例患者共接受184疗程化疗,其中2例治疗失败,血清学完全缓解率(SCR)95%。FAV组42例患者共接受190疗程化疗,其中2例治疗失败,SCR 95.2%,差异无统计学意义(P>0.05)。FPV组的1~2级腹泻、3级口腔溃疡的发生率低于FAV组,1~2级恶心呕吐、肝损害、口腔溃疡、3级中性粒细胞减少的发生率高于FAV组,差异有统计学意义(P<0.05),两组4级副反应的发生情况差异无统计学意义(P>0.05)。结论:FPV与FAV方案的疗效相当,主要副反应为胃肠道反应和骨髓毒性,微量泵持续泵入的方法减少5-FU化疗副反应的发生率,提高了患者的耐受性。FPV方案临床效果肯定,有进一步推广的价值。     

胎盘部位滋养细胞肿瘤的临床特点

1976年 Kurman 等描述了一种变异的滋养细胞疾病并称之为滋养细胞假瘤。在形态学上细胞类似滋养细胞并且侵入周围组织。对这些侵入的细胞进行免疫酶学研究提示存在有绒毛膜促性腺激素(hcG)。本病也有一些特点不同于绒癌。该肿瘤缺少通常如绒癌那样同时具有细胞滋养细胞及合体滋养细胞的两种细胞类型,在瘤体中亦无广泛坏死及出血。细胞浸润的特点是细胞潜入肌束及纤维束之间。在12例此种患者中,无因本病而     

胎盘部位滋养细胞肿瘤诊治进展

<正>早在公元前400年希波克拉底报道了子宫浮肿,现在看来很有可能就是滋养细胞疾病。1976年Kurman和Scully首先定义了"滋养细胞肿瘤"(ges-     

5-FU和VPB方案对43例恶性滋养细胞肿瘤治疗的比较

目的 评价5-FU与VPB方案治疗恶性滋养细胞肿瘤的疗效及副反应。方法 回顾性分析北京大学第一医院自1991年至2001年收治的43例恶性滋养细胞肿瘤的疗效、副反应。结果28例侵袭性葡萄胎中26例(92.9％)及15例绒癌中12例(80％)临床治愈。我院39例恶性滋养叶细胞肿瘤治疗中采用5-FU方案和VPB方案,有效率均可达91.3％,但其中5-FU方案化疗副反应发生率较VPB方案高,其中主要包括严重消化道反应、发热与感染、严重骨髓抑制,且5-FU方案每疗程住院天数较VPB方案天数长(P<0.05)。结论5-FU方案和VPB方案有效率差异无显著性,5-FU方案化疗副反应发生率较VPB方案高,住院时间长,但较VPB方案价廉。     

不同浓度瘦素对人早孕期滋养细胞MMP9表达的影响

目前普遍认为绒毛外滋养细胞侵蚀子宫螺旋小动脉参与了子宫胎盘血液循环的建立过程。作为肥胖基因产物的瘦素（1eptin）在能量代谢平衡、造血功能方面发挥很重要的作用。近年研究证实瘦素在人类胎盘滋养细胞中有表达。体外研究亦发现人工培养的滋养细胞有合成瘦素的功能，且细胞滋养细胞经诱导转变为合体滋养细胞后，其合成和分泌瘦素的能力亦显著提高。MMP是降解蜕膜基质的重要酶类，且妊娠早期以基质金属蛋白酶9（matrix metalloproteinases，MMP9）较为重要。蜕膜基质降解后，     

胎盘部位滋养细胞肿瘤1例报告

1病历摘要患者38岁,孕3产1。主因人工流产术后两个月余,血HCG异常于2010-06-14入院。患者平素月经规律,停经40余天,行超声检查,提示宫内早孕,行人工流产术,诉术中刮出绒毛状组织少许,而后一直未来月经,人流术后2个月余,行超声检查提示子宫内异常回声区,伴低阻血流。     

胎盘残留与妊娠滋养细胞疾病的甄别

胎盘残留(residual placenta)是产后30分钟胎盘未完全排除而残留于宫腔内的产后并发症,它是引起产后出血、宫腔感染的主要原因[1].妊娠滋养细胞疾病(gestational trophoblastic disease,GTD)是一组来源于胎盘滋养细胞的疾病.根据良恶性进行分类,其中良性滋养细胞疾病包括完全...     

胎盘部位滋养细胞肿瘤的病例分析

胎盘部位滋养细胞肿瘤的病例分析上海医科大学妇产科医院（２０００１１）朱关珍陆惠娟胎盘部位滋养细胞肿瘤（ｐｌａｃｅｎｔａｌｓｉｔｅｔｒｏ－ｐｈｏｂｌａｓｔｉｃｔｕｍｏｒ，ＰＳＴＴ）是一种少见而独特的子宫肿瘤。按Ｖａｒｄａｒ［１］报道在英文文献迄今共有７...     

Effects of oxygen on cell turnover and expression of regulators of apoptosis in human placental trophoblast

Pre-eclampsia (PE) and intrauterine growth restriction (IUGR) are associated with aberrant cell turnover, including increased apoptosis, in placental villous trophoblast. The increased apoptosis is associated with exaggerated expression of p53, which promotes cell cycle arrest or apoptosis via downstream proteins such as p21 or Bax. These changes in apoptosis and p53 expression are purported to result from exposure to altered oxygen tension. Using a model of villous trophoblast turnover, we examined the effect of 20%, 6% and 1% ambient oxygen (O(2)) on apoptosis, necrosis, proliferation and expression of p53 and related regulators of cell turnover, compared to both fresh tissue. Altered O(2) tension exerted an effect on cell turnover in cultured term villous tissue: cytotrophoblast proliferation was increased by culture in 20% O(2) and reduced in 1% O(2) (median proliferative index: fresh tissue=0.32%, 20% O(2)=0.9%, 6% O(2)=0.28%, 1% O(2)=0.07%). Apoptosis was increased in all culture environments, but was significantly enhanced by culture in 1% O(2) (median apoptotic index: fresh tissue=0.64%, 20% O(2)=2.96%, 6% O(2)=3.81%, 1% O(2)=9.2%). Necrotic cell death was also increased by culture in 1% O(2) compared to 6% and 20% O(2). The expression of p53, p21 and Mdm2 in both cytotrophoblast and stromal cells was increased following culture in 1% O(2). There was no alteration in the expression of Bax or Bcl-2. This study provides evidence that p53 is elevated in trophoblast following exposure to hypoxia. The potential role of the p53-pathway in the control of cell turnover in villous trophoblast and the regulation of p53 by altered O(2) tension merits further investigation.     

The effect of histone deacetylase inhibition on the expression of P-glycoprotein in human placental trophoblast cell lines

IntroductionsPlacental P-glycoprotein (P-gp), encoded by ABCB1 gene in human, plays a significant role in regulating drugs' transplacental transfer rates. Investigations on placental P-gp regulation could provide more therapeutic targets for individualized and safe pharmacotherapy during pregnancy. Currently, the epigenetic control of placental P-gp is rare. This study aimed to investigate the effect of histone deacetylases (HDACs) inhibition on P-gp expression in placental trophoblast cell lines and to explore whether HDAC1/2/3 was involved in this process preliminarily.MethodsHuman placental trophoblast cell lines (Bewo and JAR) were treated with two different HDAC inhibitors-suberoylanilide hydroxamic acid (SAHA) and trichostatin A (TSA) at different concentration gradients of 0.5, 1.0, 3.0 and 5.0 μM. Cells were harvested after 24, 48, and 72 h treatment. Total HDAC activity was detected by colorimetric assay Kits. HDAC1/2/3/ABCB1 mRNA and protein expressions were determined by real-time quantitative PCR and western-blot, respectively. Pearson correlation analysis test was performed to explore the relationship between HDAC1/2/3 mRNA and ABCB1 mRNA expression.ResultsSAHA and TSA could inhibit total HDAC activity and placental HDAC1/2/3 expression both in Bewo and JAR, but displayed a transient induction of HDAC mRNA or protein level after being treated at low dosage or prolonged exposure to drugs. Discordance in HDAC mRNA and protein expression was also observed. Placental P-gp expression was significantly induced in company with HDACs inhibition. There was a significant negative linear relationship between HDAC1/2 mRNA and ABCB1 mRNA expression.ConclusionsHDACs inhibition could up-regulate placental P-gp expression in trophoblast cells, and HDAC1/2 was most likely to be involved in this process.     

Liver X receptors mediate inhibition of hCG secretion in a human placental trophoblast cell line

Liver X receptors (LXR) alpha and beta are important regulators of lipid homeostasis in liver, adipose and other tissues. However, no such information is available for the human placenta. We determined expression of both LXR alpha and beta in placental trophoblast cell lines, BeWo and JAR. Exposure of BeWo cells to a synthetic LXR agonist, T0901317, resulted in an increase in the amount of mRNA of LXR target genes, sterol regulatory element-binding protein-1 and fatty acid synthase. T0901317 also increased the synthesis of lipids. Moreover, T0901317 resulted in a reduced secretion of hCG during differentiation of these cells. Our data for the first time demonstrate a new role for LXRs in the human placenta.     

Wnt5a inhibited human trophoblast cell line HTR8/SVneo invasion: implications for early placentation and preeclampsia

Objective: Wnt5a and Wnt signaling play potential roles in human placental and fetal development. The objective of this study is to explore the role of Wnt5a in the invasion of the human trophoblast cell line HTR8/SVneo and the probable mechanism of early placentation and preeclampsia in which Wnt5a is involved.

Methods: Human first trimester villous tissues from normal pregnancies and third trimester placentas from pregnancies with or without preeclampsia (PE) were used in the detection of the expression and subcellular location of Wnt5a. The human trophoblast cell line HTR8/SVneo was treated with 0–400?ng/ml recombinant Wnt5a to investigate the role of Wnt5a in human trophoblast invasion.

Results: Human first trimester villous is accompanied by the decreased expression of Wnt5a compared with term placenta. Upregulated Wnt5a was detected in PE placenta compared with the normal control. Wnt5a inhibited the migration and invasion of HTR8/SVneo cells with decreased integrin β1, α5 and N-cadherin. Moreover, Wnt5a downregulated β-catenin in HTR8/SVneo cells.

Conclusions: These findings strongly suggest that Wnt5a inhibits the invasion of HTR8/SVneo cells. Decreased Wnt5a facilitates early placentation, whereas increased Wnt5a contributes to the pathogenesis of PE with insufficient trophoblast invasion. Aberrant Wnt5a may function by impairing Wnt non-canonical/β-catenin signaling pathway in trophoblasts.     

Evaluation of human chorionic trophoblast cells and placental macrophages as stimulators of maternal lymphocyte proliferation in vitro

Dispersed cell suspensions of human chorion membrane and placentae were obtained by enzyme digestion and the cells examined for HLA expression and for the ability to stimulate immune cell proliferation in vitro. Chorion cells with the characteristics of trophoblast were HLA-A, B, C and Ia negative following tissue digestion whereas placental cells, primarily Fc gamma R positive macrophages, were HLA-A, B, C positive and were frequently Ia positive. When chorion and placental cell suspensions were used as stimulator cells in one-way mixed cell cultures (MCC) with maternal mononuclear leukocytes (MNL) as responder cells, chorion cells were not normally stimulatory (mean stimulation index (SI), 2.7) whereas placental cells usually were (mean SI, 11.5). No evidence for active suppression by chorion cells was obtained in a group of experiments designed to detect suppressive activity. The results support the concept of the trophoblast layer as an immunologically inert barrier between the mother and the fetus.     

Identification of human chorionic gonadotropin (HCG) secreting cells and other cell types using antibody to HCG and a new monoclonal antibody (mABlu-5) in cultures of human placental villi

Summary We have identified cells which secrete human chorionic gonadotropin (HCG) of cultures if first trimester placental villi. As a first step, we identified epithelial cells using a new monoclonal antibody. We then added HCG antibodies to the cultured cells. We found that syncytiotrophoblast (and not cytotrophoblast), Hofbauer cells and some mesenchymal cells stained with HCG antibodies.     

莉芙敏对去卵巢大鼠下丘脑视前区5-HT1AR和5-HT2AR表达的影响

目的:观察去卵巢大鼠应用黑升麻异丙醇提取物--莉芙敏(ICR)治疗1-4周后,免疫组化方法检测5-羟色胺1A受体(5-HT1AR)和5-HT2AR在大鼠下丘脑视前区表达的变化情况,为莉芙敏缓解围绝经期潮热症状的机制研究提供形态学依据.方法:雌性大鼠分为假手术组(Sham组)、去卵巢组(OVX组)、OVX后戊酸雌二醇治疗...     

Phosphorylation of JAK2 by serotonin 5-HT (2A) receptor activates both STAT3 and ERK1/2 pathways and increases growth of JEG-3 human placental choriocarcinoma cell

Serotonin 5-HT_2A receptor activation improves viability, increases DNA synthesis and activates JAK2-STAT3 and MEK1/2-ERK1/2 signalling pathways in JEG-3 human trophoblast choriocarcinoma cells. The goal of this study was to characterize the signal transduction cascade involved in 5-HT_2A receptor-induced growth of JEG-3 cells. Selective 5-HT_2A receptor agonist, DOI, induced JEG-3 cell growth was inhibited by the inhibitor of JAK2 (AG490), MEK1/2 (U0126), phospholipase C-β (PLC-β; U73122) and protein kinase C-β (PKC-β; Gö6976)), whereas the selective phosphatidylinositol 3-kinase (PI3K) inhibitor (LY294002) had no effect. Specific inhibitors of PLC-β, PKC-β and Ras (farnesylthiosalicylic acid) inhibit activation of ERK1/2, whereas the PKC-ζ inhibitor GF109203X had no effect. Interestingly, inhibition of JAK2 prevented DOI-induced phosphorylation of ERK1/2 whereas inhibition of ERK1/2 pathway had no effect on DOI-induced activation of STAT3. Taken together, our results demonstrate that both the JAK2-STAT3 and PLC-β-PKC-β-Ras-ERK1/2 signalling pathways are involved in the stimulation of JEG-3 cell growth mediated by DOI. Moreover, this study shows that activation of JAK2 by the 5-HT_2A receptor is essential to activate both STAT3 and ERK1/2 signalling pathways as well as to increase JEG-3 choriocarcinoma cell growth and survival.