TT

€340.00*

Prices excl. VAT plus shipping costs

Product number: 305027

Description	TT cells continuously produce high levels of calcitonin and CEA.Immunoreactive calcitonin was found to be produced in cell culture at levels of 3900 pg/million cells and 7700 pg/million cells 24 and 72 hours respectively, after a medium change.CEA was found to accumulate to greater than 27 ng/million cells over a 72 hours period.Chromosomal analysis of the cell line and tumors induced in nude mice reveal an aneuploid human karyotype with several marker chromosomes.The initial characterization studies of the TT cell line were conducted using early passage TT cells cultivated in RPMI 1640 medium supplemented with 15% fetal bovine serum and 1mM L-glutamine.It is not known if the neuropeptides reported to be produced by this cell line when it was grown in RPMI 1640 medium are also produced by the cells when they are cultured in Ham's F-12K medium.Chromosomal analysis of the cell line and tumors induced in nude mice reveal an aneuploid human karyotype with several marker chromosomes.
Organism	Human
Tissue	Thyroid, medulla
Disease	Hereditary thyroid gland medullary carcinoma, Multiple endocrine neoplasia type 2
Synonyms	MTC-TT

Age	77 years
Gender	Female
Ethnicity	European
Morphology	Epithelial
Growth properties	Adherent

Identifiers / Biosafety / Citation

Citation	TT (CLS catalog number 305027)
Biosafety level	1

Expression / Mutation / Susceptibility

Protein expression	Calcitonin, Carcinoembryonic Antigen(CEA)
Tumorigenic	Yes

Culture Medium	Ham's F12K
Medium supplements	10% FBS, 2.0 mM L-glutamin, 1.25 g/L L-glucose, 2.5 g/L NaHCO3
Passaging solution	Accutase
Subculturing	Remove medium and rinse the adherent cells using PBS without calcium and magnesium (3-5 ml PBS for T25, 5-10 ml for T75 cell culture flasks). Add Accutase (1-2 ml per T25, 2.5 ml per T75 cell culture flask), the cell sheet must be covered completely. Incubate at ambient temperature for 8-10 minutes. Carefully resuspend the cells with medium (10 ml), centrifuge for 3 min at 300 g, resuspend cells in fresh medium and dispense into new flasks which contain fresh medium.
Split ratio	1:2 to 1:4
Fluid renewal	2 to 3 times per week
Freeze medium	CM-1 (CLS catalog number 800100) or CM-ACF (CLS catalog number 806100)
Handling of cryopreserved cultures	The cells come deep-frozen shipped on dry ice. Please make sure that the vial is still frozen. If immediate culturing is not intended, the cryovial must be stored below -150 degree Celsius after arrival. If immediate culturing is intended, please follow the below instructions: Quickly thaw by rapid agitation in a 37 degree Celsius water bath within 40-60 seconds. The water bath should have clean water containing an antimicrobial agent. As soon as the sample has thawed, remove the cryovial from the water bath. A small ice clump should still remain and the vial should still be cold. From now on, all operations should be carried out under aseptic conditions. Transfer the cryovial to a sterile flow cabinet and wipe with 70% alcohol. Carefully open the vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of culture medium (room temperature). Resuspend the cells carefully. Centrifuge at 300 x g for 3 min and discard the supernatant. The centrifugation step may be omitted, but in this case the remains of the freeze medium have to be removed 24 hours later. Resuspend the cells carefully in 10 ml fresh cell culture medium and transfer them into two T25 cell culture flasks. All further steps are described in the subculture section.
Handling of proliferating cultures	One or two cell culture flasks come filled with cell culture medium. Collect the entire medium in 1 or 2 x 50 ml centrifuge tubes, respectively. Carefully add 5 ml of cell culture medium to each T25 cell culture flask. Control the cell morphology and confluency under the microscope. Incubate at 37 degree Celsius for a minimum of 24 hours. Spin down the collected medium at 300 x g for 3 minutes to collect the cells which may have detached during transit. If a cell pellet is visible, resuspend the cells in 5 ml of cell culture medium and transfer to a T25 cell culture flask. Incubate at 37 degree Celsius for a minimum of 24 hours.

Quality control / Genetic profile / HLA

Sterility	Mycoplasma contamination was excluded through PCR-based and luminescence-based mycoplasma assays. Bacterial or fungal contaminations are detected through daily visual cell monitoring.
STR profile	Amelogenin: X CSF1PO: 10,13 D13S317: 11 D16S539: 12,13 D5S818: 12,13 D7S820: 10,12 TH01: 6,9 TPOX: 8,11 vWA: 16,18 D3S1358: 15 D21S11: 29,32.2 D18S51: 12 Penta E: 7,13 Penta D: 13,13 D8S1179: 15,16 FGA: 21,25 D6S1043: 12,13 D2S1338: 17,23 D12S391: 15,21 D19S433: 14,15

Contamination-free cells

To identify mycoplasma contaminations we perform PCR-based and luminescence-based mycoplasma assays. We further determine any bacterial or fungal contamination through our standardized manufacturing processes.

Bulk order

Besides genomic DNA, RNA, cell pellets, and cell lysates in bulk, we can offer large quantities of assay-ready cells, plated cells in multiple formats, and frozen or growing cells. Contact us to receive a quote.

Authenticated cells

Each manufactured batch of cell lines* is authenticated via STR analysis. Contact us if you require a publication-ready STR report for your cells (*human, hamster, mouse, rat, and dog cells).

HLA alleles

HLA characterization is available from more than 200 cell lines. HLA class I -A, B, C, and Class II HLA-DPA1, -DPB1, -DQA1, DQB1, and DRB1 alleles were obtained by next-generation sequencing methodologies (NGS) for class I and class II alleles.