Carcinoma associated fibroblasts (CAFs) have recently been implicated in important aspects of epithelial solid tumor biology such as neoplastic progression, tumor growth, angiogenesis, and metastasis.
Product is for Research use only.
Frozen Vials are shipped in a Dry Ice Package.
HANDLING OF ARRIVING CELLS
Human pancreatic cancer-associated fibroblasts (CAFs) are a type of stellate cells that are found in the pancreatic tumor microenvironment. These cells play a critical role in the progression of pancreatic cancer by promoting tumor growth, invasion, and metastasis.
Pancreatic CAFs are activated by the tumor microenvironment and undergo phenotypic changes that result in increased production of extracellular matrix (ECM) proteins, growth factors, and cytokines. This leads to the deposition of a dense, fibrotic ECM that can contribute to tumor progression by enhancing tumor cell survival, promoting angiogenesis, and inhibiting immune cell infiltration.
In addition to their role in ECM remodeling, pancreatic CAFs can also promote cancer progression through other mechanisms. For example, these cells can secrete factors that promote tumor cell proliferation and survival, such as insulin-like growth factor (IGF) and vascular endothelial growth factor (VEGF). Pancreatic CAFs can also secrete factors that suppress the immune response, such as interleukin-6 (IL-6) and transforming growth factor-beta (TGF-β), which can contribute to tumor immune evasion, Regeneration Biology.
Targeting pancreatic CAFs has emerged as a potential therapeutic strategy for pancreatic cancer. Several preclinical studies have shown that targeting CAFs can reduce tumor growth and improve the efficacy of chemotherapy and immunotherapy. However, developing effective therapies that selectively target CAFs without causing toxicity to normal tissues remains a major challenge.
PROTOCOL FOR THAWING THE CELLS
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. Decontaminate by dipping in or spraying with 70% ethanol. All the operations from this point on should be carried out under strict aseptic conditions.
Transfer the vial contents to a centrifuge tube containing9.0 ml complete low serum culture medium (cat. CAFM_RG601) and centrifuge the cell suspension at approximately 125 x g for 5 to 10 minutes. Discard the supernatant.
Resuspend the cell pellet in the complete low serum culture medium at the dilution ratio recommended in the specific batch information Transfer to a 75 cm2 tissue culture flask. It is important to avoid excessive.
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alkalinity of the medium during 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).
Note: If you have any questions or need clarification regarding the protocol for culturing these cells, please reach out to email email@example.com your questions before beginning.
Note: Volumes are given for a 75 cm2 flask. Increase or decrease the amount of medium needed proportionally for culture vessels of other sizes.
Remove and discard culture medium.
Briefly rinse the cell layer with 1x PBS, remove and discard 1x PBS.
Add 2.0 to 3.0 mL of Cell Detachment solution (cat. ADF001) to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
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.
Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
To remove cell detachment solution, transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant and resuspend cells in fresh serum-free growth medium. Add appropriate aliquots of cell suspension to new culture vessels.
Place culture vessels in incubators at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:6 is recommended.