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Keratinocyte serum free medium

Manufactured by PromoCell
Sourced in Germany

Keratinocyte serum-free medium is a specialized cell culture medium designed for the growth and maintenance of human keratinocytes. It provides essential nutrients and growth factors required for the optimal proliferation and differentiation of keratinocytes in an in vitro setting, without the need for serum supplementation.

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2 protocols using keratinocyte serum free medium

1

Co-culture of NHEKs with Psoriatic DMSCs

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Total of 1×105 adult pooled NHEK cells (Promo Cell, Heidelberg, Germany) were cultured in keratinocyte serum-free medium (Promo Cell) with epidermal growth factor plus bovine pituitary extract (Gibco, Gran Island, NY) and 1% antibiotics, and seeded at 1×104 cells/ml in 25 cm2 tissue culture flasks at 37°C in 5% CO2. The medium was changed daily until the cells were confluent. NHEKs were detached using 0.25% trypsin and 0.04% EDTA. Total of 3×105 NHEKs (2×105 cells/ml) were seeded onto Transwell plates at passages 2-4 to co-culture with 1×105 DMSCs from 12 psoriasis and 12 control (500μl cell suspension at a concentration of 2×105 cells/ml), respectively. NHEKs were detached using 0.25% trypsin-0.04% EDTA after 72 h of co-culture. Afterward, NHEKs were harvested for further analyses. NHEKs cultured without any DMSCs for 72h alone served as “Untreated NHEKs”. The NHEKs co-cultured with psoriatic DMSCs and control DMSCs were labelled as “P-DMSC-Treated NHEKs” and “C-DMSC-Treated NHEKs”, respectively.
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2

Modulation of Human Hair Follicle Cells

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Human follicle dermal papilla cells (HFDPCs) (PromoCell GmbH, Heidelberg, Germany) were cultured in keratinocyte serum-free medium (PromoCell GmbH) with supplied supplement and 100 mg/mL penicillin and streptomycin (Gibco, Grand Island, NY, USA) in a 5% CO2 incubator at 37°C. Cells were used within ten passages for the experiment. The HFDPCs (1.5 × 105 cells/mL) were seeded in collagen type-I coated 96-well and 6-well plates and incubated overnight. The cells were treated with DHT (10 nM, Sigma-Aldrich, St. Louis, MO, USA) with or without finasteride (1 μM, Sigma, USA) or 50 μg/mL of S. hexaphylla extract for 24 and 72 h. Cellular morphology was observed at 200 magnification using a phase contrast microscope (Olympus, Japan).
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