Human Dermal Fibroblast - Adult (HDF-Ad)
|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.|
Identifiers / Biosafety / Citation
|Citation||Normal Human Dermal Fibroblast (Cytion catalog number 300606)|
Expression / Mutation
|Protein expression||Positive: CD73/CD90/CD105 Negative: CD14/CD34/CD45/HLA-DR|
|Viruses||Negative for: HIV-1/2, HBV, HCV, HSV1/2, CMV, EBV, HHV6, Treponema pallidum, Toxoplasma gondii, Chlamydia trachomatis, Ureaplasma urealyticum, Ureoplasma parvum|
|Culture Medium||MEM without ribonucleosides and deoxyribonucleosides|
|Medium supplements||10% FBS, 2 ng/ml hr-bFGF, 2 mM stable L-glutamine|
|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||The cells come deep-frozen shipped on dry ice. Please make sure that the vial is still frozen. If immediate culturing is not intended, the cryovial must be stored below -150 degree Celsius after arrival. If immediate culturing is intended, please follow the below instructions: Quickly thaw by rapid agitation in a 37 degree Celsius water bath within 40-60 seconds. The water bath should have clean water containing an antimicrobial agent. As soon as the sample has thawed, remove the cryovial from the water bath. A small ice clump should still remain and the vial should still be cold. From now on, all operations should be carried out under aseptic conditions. Transfer the cryovial to a sterile flow cabinet and wipe with 70% alcohol. Carefully open the vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of culture medium (room temperature). Resuspend the cells carefully. Centrifuge at 300 x g for 3 min and discard the supernatant. The centrifugation step may be omitted, but in this case the remains of the freeze medium have to be removed 24 hours later. Resuspend the cells carefully in 10 ml fresh cell culture medium and transfer them into two T25 cell culture flasks. All further steps are described in the subculture section.|
|Handling of proliferating cultures||One or two cell culture flasks come filled with cell culture medium. Collect the entire medium in 1 or 2 x 50 ml centrifuge tubes, respectively. Carefully add 5 ml of cell culture medium to each T25 cell culture flask. Control the cell morphology and confluency under the microscope. Incubate at 37 degree Celsius for a minimum of 24 hours. Spin down the collected medium at 300 x g for 3 minutes to collect the cells which may have detached during transit. If a cell pellet is visible, resuspend the cells in 5 ml of cell culture medium and transfer to a T25 cell culture flask. Incubate at 37 degree Celsius for a minimum of 24 hours.|
Quality control / Genetic profile / HLA
|Sterility||Mycoplasma contaminations are ruled out through PCR-based and luminescence-based mycoplasma assays. The absence of bacterial, fungal or yeast contamination is controlled through daily visual cell monitoring.|
To identify mycoplasma contaminations we perform PCR-based and luminescence-based mycoplasma assays. We further determine any bacterial or fungal contamination through our standardized manufacturing processes.
Besides genomic DNA, RNA, cell pellets, and cell lysates, we can offer large quantities of assay-ready cells, plated cells in multiple formats, and frozen or growing cells. Contact us to receive a quote.
Each manufactured batch of cell lines* is authenticated via STR analysis. Contact us if you require a publication-ready STR report for your cells (*human, hamster, mouse, rat, and dog cells).
HLA characterization is available from more than 200 cell lines. HLA class I -A, B, C, and Class II HLA-DPA1, -DPB1, -DQA1, DQB1, and DRB1 alleles were obtained by next-generation sequencing methodologies (NGS) for class I and class II alleles.