Human Dermal Fibroblast - Adult (HDF-Ad)

Categories: Fibroblasts Description: Human Dermal Fibroblasts, Adult (HDF-Ad), are primary cells isolated from the dermis layer of adult human skin. These cells play a crucial role in skin physiology, being responsible for the production of extracellular matrix components, including collagen and elastin, which are essential for maintaining skin structure and function. HDF-Ad cells are frequently utilized in research related to wound healing, aging, and tissue engineering, given their significant role in skin repair and regeneration processes. Additionally, they serve as an important model for studying fibroblast behavior in various dermatological conditions and diseases. HDF-Ad cells are highly responsive to external stimuli, making them a valuable tool for investigating the cellular responses to different environmental factors such as UV radiation, oxidative stress, and various pharmaceutical compounds. Their ability to proliferate and produce essential proteins under controlled conditions also makes them suitable for studies in drug development, particularly in the context of dermal toxicity and efficacy testing. These cells retain many of the physiological characteristics of their tissue of origin, providing a relevant model for in vitro studies aimed at understanding skin biology at the molecular and cellular levels. Organism: Human Tissue: Dermis Ethnicity: Caucasian Growth Properties: Adherent Citation: Human Dermal Fibroblast, Adult (HDF-Ad) (Cytion catalog number 300606) Biosafety Level: 1 Ncbi_ Taxid: 9606.0 Protein Expression: Positive: CD73/CD90/CD105 Negative: CD14/CD34/CD45/HLA-DR Tumorigenic: No Viruses: Negative for: HIV-1/2, HBV, HCV, HSV1/2, CMV, EBV, HHV6, Treponema pallidum, Toxoplasma gondii, Chlamydia trachomatis, Ureaplasma urealyticum, Ureoplasma parvum Culture Medium: MEM, w/o ribonucleosides, w/o deoxyribonucleosides (We do not supply this product, please consider other suppliers. Please let us know if you need further assistance) Supplements: Supplement the medium with 10% FBS, 2 ng/mL hr-bFGF, 2 mM stable L-glutamine Dissociation Reagent: Trypsin-EDTA Subculturing: For routine adherent cell culture: Aspirate the old culture medium from the adherent cells, and wash them with PBS to remove any remaining medium. After aspirating the PBS, add the appropriate volume of Trypsin/EDTA solution based on the culture vessel size (e.g., 1 ml for a T25 flask, 3 ml for a T75 flask) and incubate at room temperature or 37°C until the cells detach (5-10 minutes). Monitor detachment under a microscope, and gently tap the vessel if necessary to release the cells. Once detached, add complete medium to inactivate the Trypsin/EDTA, gently resuspend the cells, and transfer an aliquot of the cell suspension into a new culture vessel containing fresh medium. Place the vessel in an incubator set to 37°C with 5% CO2, and change the medium every 2-3 days. Seeding Density: 1 to 3*103 cells/cm2 Fluid Renewal: 2 to 3 times per week Freeze Medium: As a cryopreservation medium, use 90% FBS + 10% DMSO to maintain viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress. Thawing And Culturing Cells: Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit. 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. 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. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening. 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. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium. 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. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes. Sterility: 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. Safety Precautions: When planning to store a cryovial in liquid nitrogen for future thawing, it is mandatory to adhere to stringent safety measures. Appropriate protective gloves and clothing are essential, and the use of a face mask or safety goggles is required during the transfer of frozen samples to or from the liquid nitrogen tank. This is to mitigate the risk of injury from potential cryovial explosions upon removal, which can result in the projection of sharp fragments. Warranty: We stand by the promise of delivering products with high cell viability and robust culture performance. To achieve the best results, please make sure you follow the storage and culture instructions detailed in the product information sheet closely. Your adherence to these guidelines is key to success. Subject To Material Transfer Agreements: If you intend to use Cytion cell lines solely for internal research at a single research site, please complete and sign our Material Transfer Agreement (MTA) and submit it along with your order.For any commercial applications - including but not limited to fee-for-service work, quality control testing, product release, diagnostic use, or regulatory studies - please complete the Intended Use Form so we can prepare a suitable agreement tailored to your project.Please note: The MTA applies only to certain cell lines. If this notice and the MTA document appear on a product page, the agreement is applicable. For cell lines not covered by the MTA, no reference to the agreement will be shown. The MTA is not valid for customers in the Americas, China, or Taiwan. Please contact our U.S. entity to receive the appropriate agreement. Required Product 3: 860015.0 Required Product 4: 830100.0