Genome-wide gene expression analysis of neurofibroma-derived NF1+/- and NF1-/- Schwann cells

Analysis of the effect of NF1 second hit mutation to the reading of the whole human genome by comparing the gene expression profiles of neurofibroma derived Schwann cell cultures representing two different NF1 genotypes (NF1+/- and NF1-/-). Schwann cell cultures representing two different NF1 genotypes (NF1+/- and NF1-/-) were established from a total of nine tumors from three different NF1 patients (E8, E27 and S34 corresponding patients 1, 2 and 3 respectively). Total RNA was isolated from cells in passage 4. Furthermore, NF1+/- cells from 6 tumors (from patients E27 and S34) were passaged into fifth passage to test the effect of growth factor forskolin which is the separating factor between NF1+/- and NF1-/- Schwann cell culture conditions. TotalRNA samples from each culture were harvested in passage 5 (referred as Forskolin sample). For the remaining cultures, the medium was changed into proliferation medium without forskolin and after 24 hours, totalRNA samples from each culture were collected (woForskolin samples). The proliferation medium containing forskolin was given back to the remaining cultures, and after 24 hours total RNA samples from each culture were collected (ForskolinBack samples).