The term basal plate of the human placenta as a source of functional extravillous trophoblast cells.
The term basal plate of the human placenta as a source of functional extravillous trophoblast cells.
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DOI：
10.1186/1477-7827-12-7
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发表时间：
2014-01-28
期刊：
影响因子：
--
通讯作者：
Bevilacqua E
中科院分区：
文献类型：
--
作者：
Borbely AU;Sandri S;Fernandes IR;Prado KM;Cardoso EC;Correa-Silva S;Albuquerque R;Knöfler M;Beltrão-Braga P;Campa A;Bevilacqua E
绒毛外滋养层（EVT）细胞在人类胚胎着床和母胎界面的稳态中起着至关重要的作用。EVT对子宫内膜的侵袭有助于胎盘锚定、螺旋动脉重塑、免疫防御、耐受性反应以及参与建立和维持成功妊娠的几种协作性交叉作用。我们在这里报告了一个改进的协议，从人类足月胎盘的基板完全分化的EVT细胞的分离。在酶消化之前，将基板从绒毛组织和绒毛膜小心地解剖。使用30和60%Percoll梯度分离足月基底EVT细胞。一组标记物和分离细胞的特征用于确认该方法的特异性和有效性，从而可以确定它们作为胎盘研究的调查工具的潜力。分离的细胞对细胞角蛋白-7（CK-7）、胎盘生长因子、胎盘碱性磷酸酶、人类白细胞抗原G1（HLA-G1）以及α1和α5整合素呈免疫反应性，与妊娠早期胎盘绒毛的EVT标志物相似。约95%的分离细胞CK-7和82%的HLA-G1标记阳性。通过碘化丙啶掺入和台盼蓝试验排除，在EVT培养48小时期间未观察到活力的显著变化。金属蛋白酶MMP-2和MMP 9（滋养层侵袭的正调节因子）的基因表达培养48小时，也是分离的细胞的明胶分解活性。转化生长因子（TGF）-β，抑制增殖，迁移和早期EVT细胞的侵袭，也减少了transwell试验中孤立的长期EVT细胞的侵袭，而表皮生长因子是一个积极的调制器。足月基板可能是功能性EVT细胞的可行来源，其是绒毛膜衍生EVT细胞和细胞系的替代物。孤立的长期EVT细胞可能是特别有用的病理妊娠，其中绒毛外滋养层细胞的精确调节和相互作用能力受损的滋养层细胞的作用的调查。
Extravillous trophoblast (EVT) cells are of pivotal importance in human embryo implantation and homeostasis of the maternal fetal interface. Invasion of the endometrium by EVT contributes to placental anchorage, spiral artery remodeling, immunological defense, tolerogenic responses, and several collaborative cross talks involved in establishing and maintaining a successful pregnancy. We report here an improved protocol for the isolation of fully differentiated EVT cells from the basal plate of the human term placenta. The basal plate was carefully dissected from the villous tissue and the amniochorion membrane prior to enzymatic digestion. Term basal EVT cells were isolated using a 30 and 60% Percoll gradient. A panel of markers and characteristics of the isolated cells were used to confirm the specificity and efficiency of the method so that their potential as an investigative tool for placental research could be ascertained. Isolated cells were immunoreactive for cytokeratin-7 (CK-7), placental growth factor, placental alkaline phosphatase, human leukocyte antigen G1 (HLA-G1), and α1 and α5 integrins, similarly to the EVT markers from first trimester placental villi. Around 95% of the isolated cells labeled positively for CK-7 and 82% for HLA-G1. No significant change in viability was observed during 48 h of EVT culture as indicated by propidium iodide incorporation and trypan blue test exclusion. Genes for metalloproteinases MMP-2 and MMP9 (positive regulators of trophoblast invasiveness) were expressed up to 48 h of culturing, as also the gelatinolytic activity of the isolated cells. Transforming growth factor (TGF)-beta, which inhibits proliferation, migration, and invasiveness of first-trimester EVT cells, also reduced invasion of isolated term EVT cells in transwell assays, whereas epidermal growth factor was a positive modulator. Term basal plate may be a viable source of functional EVT cells that is an alternative to villous explant-derived EVT cells and cell lines. Isolated term EVT cells may be particularly useful in investigation of the role of trophoblast cells in pathological gestations, in which the precise regulation and interactive ability of extravillous trophoblast has been impaired.
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影响因子：
3.8
作者：
APLIN, JD
通讯作者：
APLIN, JD
DOI：
10.1083/jcb.109.2.891
发表时间：
1989-08
期刊：
The Journal of cell biology
影响因子：
--
作者：
Fisher SJ;Cui TY;Zhang L;Hartman L;Grahl K;Zhang GY;Tarpey J;Damsky CH
通讯作者：
Damsky CH
DOI：
10.1073/pnas.1204991109
发表时间：
2012-07-31
影响因子：
11.1
作者：
Manka, Szymon W.;Carafoli, Federico;Nagase, Hideaki
通讯作者：
Nagase, Hideaki
影响因子：
3.6
作者：
Aplin, JD;Haigh, T;Vicovac, L
通讯作者：
Vicovac, L
影响因子：
3.2
作者：
Leitner, K;Szlauer, R;Fuchs, R
通讯作者：
Fuchs, R