U-CH1

The U-CH1 cell line is the first established permanent human chordoma cell model, derived from a recurrent sacral chordoma. Chordomas are rare, slow-growing, locally invasive tumors that originate from notochordal remnants and primarily occur along the axial skeleton. U-CH1 exhibits cytogenetic features characteristic of chordoma, including clonal chromosomal aberrations such as der(1)t(1;22), deletions on chromosomes 4, 5, 6, 9, 10, and 20, and a derivative chromosome 20 resulting from t(10;20). Comparative genomic hybridization revealed recurrent DNA copy number changes in chordomas, particularly losses on 1p and 3p and gains on 7q, 5q, 12q, and 20. The cytogenetic profile of U-CH1 closely mirrors that of its parental tumor, reinforcing its biological relevance.

Functionally and molecularly, U-CH1 and other chordoma cell lines display hallmark features of chordoma, including expression of brachyury, a transcription factor considered a key diagnostic marker. U-CH1 also harbors deletions of CDKN2A and lacks p16 protein expression, a recurrent genetic alteration in chordomas. This alteration leads to hyperactivation of the CDK4/6 pathway, rendering U-CH1 sensitive to CDK4/6 inhibitors such as palbociclib. Treatment with palbociclib significantly reduced phosphorylated Rb levels and inhibited proliferation in vitro, indicating that U-CH1 can be a valuable preclinical model for evaluating cell cycle-targeted therapies. The cell line has also been validated through mRNA and protein profiling, confirming its representativeness of primary chordoma tumors in expression and genomic patterns.

37°C, 5% CO₂, humidified atmosphere.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods.

To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.