| Field | Specification |
|---|---|
| Species |
HTR-8/SVneo is a human trophoblast cell line derived from the chorionic villi of a first-trimester placenta, specifically from a 6-to-12-week-old embryo. These cells were immortalized by transfecting them with the gene encoding the simian virus 40 (SV40) large T antigen, which extends their lifespan while maintaining characteristics typical of extravillous invasive trophoblasts. This cell line expresses several key markers associated with extravillous trophoblasts, including insulin-like growth factor II (IGF-II), NDOG-5, proliferating cell nuclear antigen (PCNA), and a range of integrins (α1, α3, α5, αv, and β1 subunits, along with the αvβ3/β5 vitronectin receptor). It is negative for macrophage marker 63/D3, endothelial cell marker factor VIII, and α6 and β4 integrin subunits, confirming its trophoblast lineage and distinguishing it from other cell types such as macrophages and endothelial cells.
HTR-8/SVneo cells are widely used as a model to study trophoblast invasion and placental biology, particularly the epithelial-to-mesenchymal transition (EMT), which is crucial for trophoblasts' invasive behavior during placental development. Research has shown that these cells exhibit a mixed population of epithelial and mesenchymal phenotypes, with the ability to undergo EMT under standard culture conditions. This transition is mediated by TGF-β signaling, which promotes the mesenchymal phenotype, as evidenced by the upregulation of mesenchymal markers such as vimentin and the downregulation of epithelial markers like E-cadherin. This makes HTR-8/SVneo a valuable in vitro model for studying the molecular mechanisms underlying EMT in trophoblasts and its implications in both normal placental development and pregnancy-related disorders.
Studies have further demonstrated that HTR-8/SVneo cells can form spheroids, which predominantly express epithelial markers. When these spheroids are re-plated in 2D culture, the cells exhibit a shift towards a mesenchymal phenotype, indicating an ongoing EMT process. This cell line's unique properties, including its responsiveness to TGF-β and its mixed epithelial-mesenchymal nature, provide critical insights into the complex cellular dynamics of trophoblast invasion and the regulation of placental development, offering a robust platform for investigating pregnancy-related pathologies such as pre-eclampsia and intrauterine growth restriction.
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Viruses: Simian virus 40 (transfected with pSV3neo plasmid containing the early region of SV40)
- cultureMedium: RPMI 1640, w: 2.0 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a)
- supplements: Supplement the medium with 10% FBS
- dissociationReagent: Accutase
- subculturing: Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
- freezeMedium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
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