Tau RD P301S FRET Biosensor cell line生物传感器细胞株 BioVector NTCC质粒载体菌种细胞基因保藏中心

Tau RD P301S FRET Biosensor cell line生物传感器细胞株

Organism Homo sapiens, human Tissue embryonic kidney Product Format frozen 1.0 mL Morphology epithelial Culture Properties adherent Biosafety Level 2 [Cells contain SV40 and CMV viral DNA sequences] Age fetus Gender female Applications This cell line has been engineered to report tau seeding activity. Tau seeds introduced into the culture media of Tau RD P301S FRET Biosensor Cells can nucleate the aggregation of the endogenous tau reporter proteins. This aggregation produces a FRET signal which can be measured via microscopy, microplate readers or flow cytometry. Image Derivation The Tau RD P301S FRET Biosensor cells were derived by transducing HEK293T cells with 2 separate lentivirus constructs encoding tau RD P301S-CFP and tau RD P301S-YFP. Dual-positive cells were identified by FAC sorting and were cloned and isolated using cloning cylinders. Complete Growth Medium The base medium for this cell line is BioVector-formulated Dulbecco's Modified Eagle's Medium. To make the complete growth medium, add the following components to the base medium: 2mM L-alanyl-L-glutamine fetal bovine serum to a final concentration of 10% Subculturing Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Remove and discard culture medium. Add 2.0 to 3.0 ml of 0.25% (w/v) Trypsin - 0.53 mM EDTA (BioVector 30-2101) solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes). Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels. Incubate cultures at 37°C. Subcultivation Ratio: 1:6 to 1:12 is recommended Cryopreservation Freeze Medium: Fetal Bovine Serum, 90%; DMSO, 10% Storage Temperature: liquid nitrogen vapor phase Culture Conditions Temperature: 37°C Atmosphere: air, 95%; carbon dioxide (CO2), 5% Volume 1.0 mL STR Profile Amelogenin: X CSF1PO: 11, 12 D13S317: 12, 14 D16S539: 9, 13 D5S818: 8, 9 D7S820: 11 TH01: 7, 9.3 TPOX: 11 vWA: 16, 19 COI Human Year of Origin 2014 References Holmes BB, et al. Proteopathic tau seeding predicts tauopathy in vivo. Proc. Natl Acad. Sci. 111(41): E4376-4385, 2014. PubMed: 25261551

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