HROC222 T1 M2

HROC222 T1 M2 is a human colorectal adenocarcinoma cell line established within the HROC (Hansestadt Rostock Colorectal Cancer) model collection from a primary tumor resected from an adult patient. The designation “T1” indicates that the specimen was obtained at the first surgical time point, while “M2” denotes the corresponding in vitro model generated from this tumor. The HROC platform integrates comprehensive biobanking, standardized molecular annotation, and parallel establishment of patient-derived xenografts (PDX) and permanent low-passage cell lines, enabling clinically annotated translational research models.

Generation of HROC222 T1 M2 followed standardized procedures involving mechanical dissociation of freshly resected tumor tissue, preparation of single-cell suspensions, and seeding onto collagen-coated culture plates in defined tumor cell culture medium supplemented with glutamine, antibiotics, and antimycotics. Across the HROC cohort, permanent primary colorectal cancer cell lines were successfully established from approximately 13% of attempted specimens. Statistical analysis identified higher tumor grading as significantly associated with successful primary cell line establishment, while advanced nodal status showed a positive trend. In multivariate analysis across the collection, nodal involvement emerged as an independent predictor of model establishment success.

The HROC collection encompasses all major molecular subtypes of colorectal carcinoma, including chromosomal instability (CIN), CpG island methylator phenotype (CIMP), microsatellite stable (MSS), and microsatellite instability-high (MSI-H) tumors, as well as diverse mutational backgrounds affecting key driver genes such as KRAS, BRAF, TP53, APC, and PIK3CA. HROC222 T1 M2 was generated within this rigorously characterized framework, permitting integration with detailed clinicopathological and molecular data and, where available, corresponding PDX material. As a low-passage, patient-derived colorectal carcinoma model, HROC222 T1 M2 is suitable for investigations of tumor biology, genotype-phenotype relationships, and preclinical therapeutic testing within precision oncology research.

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.