ATDC5

ATDC5 is a murine chondrogenic cell line derived from mouse teratocarcinoma cells and is widely used as an in vitro model for studying chondrogenesis and cartilage development. This cell line undergoes sequential chondrogenic differentiation, mimicking in vivo processes such as cellular condensation, the expression of early chondrocytic markers like type II collagen and aggrecan, and the transition to hypertrophic chondrocytes, marked by type X collagen expression and matrix mineralization. Due to its ability to proliferate and differentiate efficiently, ATDC5 serves as a valuable model for exploring molecular mechanisms related to skeletal development, especially endochondral ossification.

ATDC5 cells have been extensively used to study the influence of various growth factors, hormones, and transcription factors on chondrogenesis. For instance, transforming growth factor-beta (TGF-β) has been shown to promote early chondrogenic differentiation by modulating the expression of extracellular matrix components like fibronectin. Similarly, bone morphogenetic proteins (BMPs), particularly BMP-2, -4, and -7, play a critical role in promoting different stages of chondrocyte differentiation in ATDC5. Moreover, the activation of transient receptor potential vanilloid 4 (TRPV4) channels in these cells, combined with hyaluronan, has been demonstrated to enhance the expression of key chondrogenic markers such as SOX9 and Aggrecan, further supporting their utility in cartilage tissue engineering studies.

This cell line has been instrumental in proteomics research as well, showing that ATDC5 cells can synthesize major cartilage extracellular matrix (ECM) components like aggrecan and type II collagen, along with the proper post-translational modifications required for cartilage function. Its capacity to recapitulate crucial ECM biosynthesis events makes ATDC5 an indispensable model for studying cartilage formation and related pathologies.

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.