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Imunoanalýza
Immortalized Rat Embryonic Fibroblasts (18IM)
| Unit |
1×10⁶ cells / 1.0 ml |
|---|---|
| Cell Type |
Immortalized Cells |
| Tissue Type |
Embryo |
| Organism |
Rat (R. norvegicus) |
| Donor History | |
| Growth Properties |
Adherent ,polygonal |
| Growth Conditions |
Use of PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. PriGrow V (TM015) + 5% FBS + 0.5 mg/ml G418 (G271) +1% Penicillin/Streptomycin Solution (G255) ,37.0°C ,5% CO₂ |
| Immortalization Method |
Transfection of plasmid encoded with GFP-fused S18–2 (GFP-S18–2) carrying a neomycin resistance gene |
| Full Information |
https://www.abmgood.com/immortalized-rat-embryonic-fibroblasts-18im.html |
Immortalized Rat Embryonic Fibroblasts (18IM) are derived from rat embryonic fibroblasts isolated from 13 to 14 day old fetuses which were transfected with a plasmid containing S18-2. Transfected cells were selected for with 0.5 mg/mL G418. 18IM cells display embryonic stem cell-like properties after immortalization and subsequent loss of mesodermal markers thereafter. Loss of contact inhibition was also observed and cells were also capable of showing anchorage-independent growth. Immunostaining showed the presence of stem cell markers, such as SSEA-1, and the loss of mesodermal markers, such as vimentin and smooth muscle actin. RT-PCR also confirmed the loss of mesodermal markers, FUT4 and vimentin, and an up-regulation of stem cell markers, Sox2 and Oct3/4. This cell line may be useful as a model for the role of S18-2 in other cell types and further studies in the binding partners of S18-2 in differentiated tissues. Western blot analysis for presence of S18-2 in cells.
