Immortalized Human Pancreatic Cancer-Associated Fibroblasts
Name of Products Immortalized Human Pancreatic Cancer-Associated Fibroblasts
Catalog Number CAF 3118-IMRG
Product Format Frozen Vial
APPLICATIONS: Cell Assays
TYPE: Human Primary CAFs
STORAGE: Keep frozen in liquid nitrogen until plating
SHIPPING: Frozen (Dry Ice/Liquid Nitrogen)
FORMAT A: Frozen
SPECIES REACTIVITY: Human
Size: 1,000,000 cells/vial
*Suggested Medium: CAF Growth Medium (CAF01)
Immortalized human pancreatic cancer-associated fibroblasts (CAFs) are a type of cell line derived from fibroblasts present in the pancreatic tumor microenvironment. These cells have been genetically modified to bypass senescence and have acquired the ability to grow and divide indefinitely in culture.
Pancreatic CAFs play a critical role in pancreatic cancer progression by promoting tumor growth, invasion, and metastasis. Immortalized human pancreatic CAFs can be used to study the complex interactions between fibroblasts and cancer cells in the tumor microenvironment and to investigate the mechanisms underlying fibroblast activation and function in pancreatic cancer.
Immortalized human pancreatic CAFs can be used to evaluate the effects of potential anti-cancer therapies on fibroblast activation and function. For example, these cells can be used to test the efficacy of drugs that target fibroblast activation pathways, such as TGF-β inhibitors or stromal cell-targeting agents. They can also be used to evaluate the effects of chemotherapy or radiation on the tumor microenvironment and the role of CAFs in promoting drug resistance.
In addition, immortalized human pancreatic CAFs can be used to investigate the role of specific genes or signaling pathways in fibroblast activation and function. For example, these cells can be used to study the effects of gene knockdown or overexpression on fibroblast activation and the secretion of cytokines, growth factors, and extracellular matrix components.
Overall, immortalized human pancreatic CAFs are valuable tools for studying the complex interactions between fibroblasts and cancer cells in the context of pancreatic cancer and for developing new therapeutic strategies targeting the tumor microenvironment.
GENERAL INFORMATION: hTert-Human Pancreatic CAF has been cultured/passaged after being infected with hTert-lentiviral particles under Special Formulated CAF Growth Medium. Human Pancreatic CAF is isolated from human Pancreatic 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 Pancreatic 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 Pancreatic 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
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)