Immortalized Rat Embryonic Striatal (M213-2O) Cells-tsSV40T
Cat.No.: CSC-I9297L
Species: Rattus norvegicus
Source: Straitum
Morphology: Multipolar polygonal
Culture Properties: Adherent
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Note: Never can cells be kept at -20 °C.
CIK-HT004 HT® Lenti-SV40 (tsA58 temperature sensitive mutant) Lentivirus Immortalization Kit
2) Test growth in permissive and non-permissive temperatures
Immortalized Rat Embryonic Striatal (M213-2O) Cells-tsSV40T are neuronal progenitor-like cells from embryonic striatal cells that have been made conditionally immortal through transfection with temperature-sensitive SV40 large T antigen (tsSV40T). Permissive conditions allow proliferation, while non-permissive conditions allow differentiation. tsSV40T will allow growth when cultured at a permissive temperature (typically 33°C). In this state the tsSV40T antigen is active and allows for prolonged proliferation and stable expansion of cells. At restrictive temperatures (non-permissive temperatures such as 37-39°C), SV40 T antigen function is decreased/inactivated which allows cells to arrest growth and differentiate. After differentiation cells display morphological and biochemical properties characteristic of striatal neurons such as neurite outgrowth and expression of neuronal markers such as βIII-tubulin, and microtubule associated protein.
Originating from striatal cells they may be used to study basal ganglia and striatum associated disorders including Huntington's disease, parkinsonian syndromes, and other movement disorders. In addition, this cell line can be used to examine neurotransmitter receptor signaling, calcium flux, oxidative stress, and neurotoxicity as well as screen neuroprotective agents. The cell line can also be used for functional analyses of genes within a neuronal lineage.
The LPS-Induced in Ammatory Circuit in Gabaergic M213-2O Cells Was Negatively Regulated by Mir-322 5p
MicroRNAs regulate gene expression post-transcriptionally, with some dysregulated miRNAs implicated in epileptogenesis. Zhou et al. investigated whether microRNA-322-5p modulates seizures and neuronal damage by targeting the NFκB-TLR4 inflammatory signaling pathway.
A pilocarpine-induced status epilepticus (SE) rat model was established, and hippocampal pathology was confirmed by immunohistochemistry. Next, they attempted to simulate the neuroin ammatory reactions by treating M213-2O cells with LPS. They found that LPS dose-dependently decreased the cell viability of M213-2O cells (Fig. 1A). Next, western blots of LPS-treated M213-2O cells demonstrated dose-dependent decreased expression of in ammatory markers including TRAF6, TRL4, RelA, and IL-6 (Fig. 1B). Consistently, the miR-322-5p level was signi cantly lower in LPS-treated M213-2O cells compared to their nontreated counterparts (Fig. 1C). Subsequently, they manipulated the level of miR-322-5p using antagomir (downregulation) and mimic (upregulation) molecules in M213-2O cells. Antagomir transfection led to the signi cantly increased expression of TRAF6 (its target), IRF5, and RelA, while decreased expression of GABA and GAD1 (Fig. 1D); this phenomenon was reversed in mimic-transfected M213-2O cells (Fig. 1D).
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