The following primary dermal cells and cell models are available for use in research and drug discovery.
Human Dermal Fibroblasts (HDF)
Dermal Fibroblasts play an important regulatory role in wound repair of the skin and also in dermal inflammation. If Dermal Fibroblasts are deregulated, wound repair is often excessive and this can lead to keloids and hypertrophic scars in the skin.
Tissues: Breast Skin, Abdominal Skin, Foreskin
Cell Isolation: Incubate small pieces of skin with Dispase II overnight in 4°C. Separate dermis from epidermis. Incubate dermis with Liberase TM for 4 hours at 37°C. After filtration, cultivate washed cells in Fibroblasts Growth Medium at 50 000 cells/cm².
Cryopreservation: Cryopreservation of cells at P1 (or P2) in >500 000 cells/mL density.
Culture: Seed HDF at 10 000 cells/cm² in Fibroblasts Growth Medium Use cells until passage P5.
Specific Markers: P4HB and Collagen I/III
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Human Epidermal Keratinocytes (HEK)
Keratinocytes produce a variety of cytokines, growth factors, interleukins and complement factors and are important for studying wound healing, inflammation, and immune response.
Tissues: Breast Skin, Abdominal Skin, Foreskin
Cell Isolation: Incubate small pieces of skin with Dispase II overnight in 4°C. Separate dermis from epidermis. Incubate epidermis with trypsin/EDTA for 20 minutes at 37°C. After filtration, cultivate washed cells (Melanocytes and Keratinocytes) in Melanocytes Growth Medium at 150 000 cells/cm². Separate HEM and HEK with trypsin /EDTA. Remove the HEM and let the HEM with Keratinocytes Growth Medium.
Cryopreservation: Cryopreservation of cells at P0-P1 in >500 000 cells/mL density.
Culture: Use cells until passage P2 at a density of 10 000 cells/cm2 in Keratinocytes Growth Medium.
Specific Markers: Cytokeratin 14 and Cytokeratin 1.10
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Human Epidermal Melanocytes (HEM)
Melanocytes produce the protective skin pigment, melanin and are located in the bottom layer of the skin’s epidermis. Changes in skin coloration often accompany inflammatory reactions and the number of melanocytes is increased in diseases such as psoriasis, lichen planus and dermatitis. Melanocytes may be useful in determining the reactivity of the skin towards external stimuli such as UV exposure and chemicals.
Tissues: Breast Skin, Abdominal Skin, Foreskin
Cell Isolation: Incubate small pieces of skin with Dispase II overnight in 4°C. Separate dermis from epidermis. Incubate epidermis with trypsin/EDTA for 20 minutes at 37°C. After filtration, cultivate washed cells (Melanocytes and Keratinocytes) in Melanocytes Growth Medium at 150 000 cells/cm². Separate HEM and HEK with trypsin /EDTA. Seed HEM at 106 cells/cm² in Melanocytes Growth Medium.
Cryopreservation: Cryopreservation of cells at P2 in >500 000 cells/mL density.
Culture: Use cells until passage P5 at a density of 10000cells/cm2 in Melanocytes Growth Medium.
Specific Markers: Tyrosinase and Melanocyte Surface Marker
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Human Preadipocytes (HPAd)
Adipose tissue is an endocrine organ involved in regulating physiologic and pathologic processes, including inflammation. It synthesizes and secretes hormones such as leptin and adiponectin as well as cytokines and chemokines, which can lead to a chronic sub-inflammatory state resulting in cardiovascular complications linked to obesity and insulin resistance. Pre-adipocytes differentiate into mature adipocytes by a complex mechanism involving several different stimuli. CellMade provides a pre-adipocyte differentiation cell model for investigating these important inflammatory processes.
Cell Differentiation Protocol: Add adipocyte differentiation medium to primary human adipocytes at 80% confluency. Allow differentiation for 10-14 days in a humidified 37°C, 5% CO2 incubator.
Adipocyte Differentiation Medium: DMEM, d-Biotin, Insulin, DXM,IBMX, L-Thyroxine, Ciglitazone.
Detection: OilRedO staining
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Human Dermal Microvascular Endothelial Cells (HDMVEC)
Endothelial cell system for the study of angiogenesis, tumor metastasis, wound healing, inflammation, macromolecule transport and drug metabolism.
Tissue: Foreskin
Cell Isolation: Incubate small pieces of skin with Dispase II overnight in 4°C. Separate dermis from epidermis. Incubate dermis with Collagenase Ia for 75 minutes at 37°C. After filtration, cultivate washed cells (HDMVEC and HDF) in HDMVEC Growth Medium at 50 000 cells/cm². Detach cells with Trypsin/EDTA. Make a CD31 magnetic beads separation to isolate HDMVEC from HDF. Seed HDMVEC (CD31+) at 5000 cells/cm2 in HDMVEC Growth Medium.
Cryopreservation: Cryopreservation of cells at P2 in >500 000 cells/mL density.
Culture: Use cells until passage P5 at 10 000 cells/cm2 in HDMVEC Growth Medium.
Specific Markers: CD31 and Von Willebrand Factor
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