Immortalized Human Colorectal Cancer-Associated Fibroblasts
Name of Products Immortalized Human Colorectal Cancer-Associated Fibroblasts
Catalog Number CAF 715-IMRB
Product Format Frozen Vial
Cell Number 1,000,000 cells/vial
*Suggested Medium: CAF Growth Medium (CAF01)
Immortalized human colorectal cancer-associated fibroblasts (CAFs) are a type of cell line derived from human fibroblasts that are associated with colorectal cancer (CRC) tumors. CAFs play an important role in the development and progression of CRC by supporting tumor growth, invasion, and metastasis.
Immortalized CAFs are created by introducing genetic modifications into primary CAFs that allow them to replicate indefinitely in culture, which enables researchers to study their behavior and function in vitro. These cell lines are commonly used in cancer research to investigate the interactions between tumor cells and the tumor microenvironment.
Studies have shown that CAFs can promote tumor growth and invasion by secreting growth factors, cytokines, and extracellular matrix proteins that stimulate angiogenesis and facilitate the invasion of cancer cells into surrounding tissues. CAFs may also play a role in immune evasion and resistance to chemotherapy.
Immortalized CAF cell lines are a valuable tool for studying the complex interactions between cancer cells and the tumor microenvironment in CRC and other types of cancer. They can be used to identify new therapeutic targets and develop new treatment strategies for cancer.
GENERAL INFORMATION: hTERT-Human Colorectal CAF has been cultured/passaged after being infected with hTERT-lentiviral particles under Special Formulated CAF Growth Medium Human Colorectal CAF is isolated from human Colorectal tumor tissue. Cells are grown in T75 tissue culture flasks pre-coated with AlphaBioCoat solution for 30 min and incubated in CAF Growth Medium for 3-7 days. Each vial contains at least 1,000,000 cells per ml.
Product Testing: Human Colorectal CAF is tested for negative expression of von Willebrand Factor Expression/Factor VIII, cytokeratin 18, andalpha-smoothh muscle actin. Cells are negative for bacteria, yeast, fungi, and mycoplasma. Cells express CAF markers FAP, PDGFR, Vimentin, PDPN, and CD70. Cells can be expanded for 3-5 passages at a split ratio of 1:2 or 1:3.
Laboratory Applications: Human Colorectal CAF can be used for the assay of cell-cell interaction, adhesion, PCR, Western blot, immunoprecipitation, immunofluorescent flow cytometry, or generating cell derivatives for desired research applications.
SHIPPING Frozen Vials on Dry Ice:
To ensure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in the liquid nitrogen vapor phase and not at -70°C. Storage at -70°C will result in loss of viability. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water. Thawing should be rapid (approximately 2 minutes).
Remove the vial from the water bath as soon as the contents are thawed and decontaminate by dipping in or spraying with 70% ethanol. All the operations from this point on should be carried out under strict aseptic conditions.
It is recommended that the cryoprotective agent be removed immediately. Centrifuge the cell suspension at approximately 125 x g for 5 to 10 minutes. Discard the supernatant and resuspend the cell pellet in an appropriate amount of fresh growth medium.
Add 6.0 to 8.0 mL of AlphaBioCoat to the T-Flask for 15 minutes. Aspirate the solution after 15 minutes, and rinse with 8ml of 1XPBS. Discard the 1XPBS. Transfer the cells to an appropriate size T-Flask.
It is important to avoid excessive alkalinity of the medium during the recovery of the cells. It is suggested that, prior to the addition of the vial contents, the culture vessel containing the growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6).
Incubate the culture at 37°C in a suitable incubator. A 5% CO2 in air atmosphere is recommended if using the medium described on this product sheet.
Subculturing procedure:
Volumes used in this protocol are for a 75 cm2 flask; proportionally reduce or increase the amount of dissociation medium for culture vessels of other sizes. T-75 flasks are recommended for subculturing this product.
*Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
Remove and discard the culture medium. Briefly rinse the cell layer with 1XPBS to remove all traces of serum that contains the inhibitor.
Add 2.0 to 3.0 mL of Cell Detachment solution to the flask and observe cells under an inverted microscope until the cell layer is dispersed (usually within 5 to 15 minutes).
Add 6.0 to 8.0 mL of AlphaBioCoat to the T-Flask for 15 minutes. Aspirate the solution after 15 minutes, and rinse with 8ml of 1XPBS. Discard the 1XPBS.
Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
Add appropriate aliquots of the cell suspension to new culture vessels.
Incubate cultures at 37°C. Sub-cultivation Ratio: A sub-cultivation ratio of 1:2 to 1:3 is recommended Medium Renewal: Every 3 to 4 days
Reagents for cryopreservation: Complete growth medium supplemented with 5% (v/v) DMSO. Catalog # (CGM542)
