MicroRNA profiling of human dermal fibroblasts - miR-17 and miR-19b to investigate down-regulatation in human aging

ABSTRACT: Normal human fibroblasts were isolated from skin biopsies of two different healthy causcasian patients, a 41-years old female (HDF-1) and a 16-years old male (HDF-5). HDF-5 and HDF-1 cells were cultivated in DMEM/HAM’s F-12 medium (Biochrom KG, Berlin, Germany) supplemented with 10% fetal calf serum (FCS) 4 mM L-glutamine (Sigma) and Primocin antibiotics (100 µg/mL) grown at 37 °C in ambient atmosphere containing 5% CO2. Cells were grown until confluence was reached and routinely passaged using 1:4 or 1:3 split ratios. Upon decrease in growth rate and entry of irreversible growth arrest split ratio was changed to 1:2 until cells stopped dividing and fresh medium was added at least every 7 days. HDF-1 cells reached senescence after 54 population doublings, while HDF-5 cells had stopped growing after 72.7 population doublings (see Fig. S2 for growth curves and staining of senescent associated beta-galactosidase).
Senescent HDF RNA samples were compared to young HDF RNA samples in terms of microRNA expression using Exiqon LNA microRNA microarrays.

ORGANISM(S): Homo sapiens

SUBMITTER: Matthias Hackl 

PROVIDER: E-MEXP-2386 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress