HC11

The HC11 cell line, a clone derived from the COMMA-1D parental cell line, is an epithelial cell line sourced from the mammary gland of a mid-pregnant BALB/c mouse. This particular clone was isolated through transfection and was subsequently selected for its ability to induce beta-casein protein in response to prolactin. As a model, HC11 is particularly noted for its responsiveness to prolactin and other lactogenic hormones like insulin and dexamethasone, which facilitate the production of milk proteins such as beta-casein.

In terms of cellular behavior and characteristics, HC11 cells are capable of differentiation in culture conditions that do not require the addition of a complex extracellular matrix or co-culture with other cell types. This simplifies the use of HC11 cells in various experimental setups, focusing on the cellular mechanisms of mammary gland function and development. Notably, HC11 cells autonomously produce key extracellular matrix proteins, including laminin, which are crucial for their structure and function. The gene expression profile of HC11 cells varies with their differentiation state: undifferentiated cells express genes such as Lgals1, Ran, Jam-A, Bmpr1a, Nfkbiz, Trib 1, Rps21, and ler3, while differentiated cells express Id1, highlighting the dynamic changes in gene expression associated with mammary epithelial cell differentiation.

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.