Journal: International Journal of Molecular Sciences
Article Title: Availability of mRNA Obtained from Peripheral Blood Mononuclear Cells for Testing Mutation Consequences in Dystrophic Epidermolysis Bullosa
doi: 10.3390/ijms222413369
Figure Lengend Snippet: Type VII collagen expression in cells isolated from PBMCs obtained using the Human Mesenchymal Stem Cell Enrichment Cocktail. ( a ) COL7A1 mRNA expression of cultured normal human epidermal keratinocyte (NHEK), normal human dermal fibroblast (NHDF), Peripheral blood mononuclear cell (PBMC), and human mesenchymal stem cell (hMSC) was analyzed using reverse transcription quantitative polymerase chain reaction (RT-qPCR). ( b ) The isolated cells were cultured in mesenchymal stem cell culture medium for 14 days. Scale bar, 100 μm. ( c ) COL7A1 mRNA expression in isolated cells. Cells were treated with or without 1 nM of transforming growth factor- (TGF-β). Quantitative analysis of COL7A1 mRNA expression was performed using RT-qPCR.
Article Snippet: To assess this possibility, we isolated MSCs from PBMCs using the RosetteSep Human Mesenchymal Stem Cell Enrichment Cocktail (Stem Cell Technologies) and cultured them with MesenPRO RS Medium (Thermo Fisher Scientific).
Techniques: Expressing, Isolation, Cell Culture, Reverse Transcription, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Stem Cell Culture