Human Dermal Fibroblast - Adult (HDF-Ad)
Product number:
300606
General information
Description | Primary Human Dermal Fibroblasts (HDF) are isolated from the dermis of adult foreskin and are provided in a cryopreserved format. HDF are self-renewing stromal cells that can differentiate into at least three different lineages: adipocytes, osteoblasts and chondrocytes. |
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Organism | Human |
Tissue | Dermis |
Characteristics
Ethnicity | Caucasian |
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Growth properties | Adherent |
Identifiers / Biosafety / Citation
Citation | Human Dermal Fibroblast, Adult (HDF-Ad) (Cytion catalog number 300606) |
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Biosafety level | 1 |
Expression / Mutation
Protein expression | Positive: CD73/CD90/CD105 Negative: CD14/CD34/CD45/HLA-DR |
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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 |
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) |
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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 | Human Dermal Fibroblast, Adult (HDF-Ad) cells are shipped in a deep-frozen state on dry ice. Upon receipt, confirm that the vial remains frozen. For storage, place the cryovial immediately at temperatures below -150 degrees. If you plan to culture the cells immediately, swiftly thaw the vial by shaking it in a 37 degrees water bath with clean water and an antimicrobial agent for 40-60 seconds. Remove the vial once a small ice clump persists, ensuring it remains cold. Proceed with all subsequent steps under aseptic conditions. In a sterile flow hood, disinfect the cryovial with 70% ethanol. Then, gently open the vial and transfer the cell suspension into a 15 ml centrifuge tube pre-filled with 8 ml of room temperature culture medium. Gently mix the cells. For cell separation, centrifuge at 300 x g for 3 minutes and dispose of the supernatant. Skipping centrifugation is optional, although any residual freezing medium should be removed after 24 hours. Resuspend the pellet gently in 10 ml of fresh culture medium and divide between two T25 culture flasks. Follow the subculture protocol for subsequent steps. |
Quality control / Genetic profile / HLA
Sterility | Mycoplasma contamination is rigorously 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. |
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