Human Dermal Fibroblast, Adult (HDF-Ad)

Primary Human Dermal Fibroblasts, Adult (NHDF-Ad) are isolated from the dermis of adult skin. They are used in research on skin aging, fibrosis and wound healing and as the dermal component of full-thickness skin equivalents.

Derived from adult skin, NHDF-Ad are a relevant model for age-related processes: in dermal fibroblasts, repeated UVA exposure induces MMP-1, oxidative stress and senescence markers that mirror features of photoaged dermis. They synthesize type I collagen and respond to TGF-β via SMAD2/3 signaling, the central pathway in dermal fibrosis.

37°C, 5% CO₂, saturated humidity.

Subculture the cells at about 75 % confluence. Denser cultures undergo irreversible contact inhibition and stop proliferating. Use the subculture reagents recommended on the certificate of analysis.

Post-thaw viability ≥ 70 % viable cells. More than 80 % confluence within the expected recovery period.

1. Thaw the cryovial quickly in a 37 °C water bath, swirling gently. Just before the last ice has melted, move the vial to a laminar flow hood and decontaminate its outer surface.

2. Transfer the cell suspension into a 15 ml centrifuge tube, add 10 ml of Fibroblast Growth Medium and centrifuge at 250 × g for 5 minutes.

3. Remove the supernatant and resuspend the cell pellet in 10 ml of Fibroblast Growth Medium. Check cell number and viability.

4. Divide the suspension between two 25 cm² culture flasks (5 ml each). Using two flasks reduces the risk of losing all cells if a passage fails.

5. Incubate at 37 °C, 5 % CO₂ and saturated humidity, with the screw caps opened by half a turn for gas exchange.

6. Change the medium after 24 hours to remove unattached cells, then every 48 to 72 hours, using pre-warmed medium only.

7. Subculture the cells at about 75 % confluence. Denser cultures undergo irreversible contact inhibition and stop proliferating. Use the subculture reagents recommended on the certificate of analysis.

8. Recommended seeding density: more than 4,000 cells/cm².

Mycoplasma contamination is excluded by PCR.

To ensure there is no bacterial, fungal or yeast contamination, cultures are subjected to visual inspection.

For long-term preservation, store vials in liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable for up to one week only.

Cryopreserved primary cells are shipped on dry ice. On receipt, check the cryovial for damage and transfer it without delay to appropriate storage.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.