Human Dermal Fibroblast - Adult (HDF-Ad)

General information

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

Handling

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)

Medium supplements — Supplement the medium with 10% FBS, 2 ng/mL hr-bFGF, 2 mM stable L-glutamine

Passaging solution — Trypsin-EDTA

Subculturing — Remove the culture medium and rinse the adherent cells using PBS without calcium and magnesium (3-5 ml PBS for T25, 5-10ml for T75 cell culture flasks). Add trypsin-EDTA 0.

25% solution, 1-2ml per T25, 2.5ml per T75 cell culture flask, the cell sheet must be covered completely, and incubate at 37 degree Celsius for 10 min.

Stop the trypsin activity using FBS-containing cell culture medium. Dispense into new flasks which contain fresh cell culture medium.

Seeding density — 1 to 3*10^3 cells/cm²

Fluid renewal — 2 to 3 times per week

Freeze medium — CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100)

Handling of cryopreserved cultures — 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.

Optionally, skip centrifugation but remove any remaining freezing medium after 24 hours.
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.