Journal: Molecular Therapy
Article Title: Neural Stem Cells Engineered to Express Three Therapeutic Factors Mediate Recovery from Chronic Stage CNS Autoimmunity
Figure Lengend Snippet: NT-3/IL-10/LINGO-1-Fc promotes NSC proliferation and differentiation in vitro . ( a ) NSCs transduced with NT-3/IL-10/LINGO-1-Fc were plated on poly-D-lysine/laminin coated coverslips at a density of 2.0 × 10 4 cell/ml in 24-well plates and cultured in NSC proliferation medium. After 48 hours of culture with Dox (1.5 μg/ml), cells were stained with anti-Ki67 antibody. ( b ) Proliferation of NSCs was quantified as the percentage of Ki67-positive cells in all GFP-positive cells. ( c ) Examples of NSCs that differentiated into mature oligodendrocytes (MBP + ), neurons (NF-H + ), and astrocytes (GFAP + ) in specific differentiation medium for 2 weeks. Scale bar = 100 µm. ( d ) Quantitative analysis of differentiated cells. Data are shown as mean values ± SD ( n = 5 each group) and are representative of three experiments. Groups designated by the same letter are not significantly different, while those with different letters (A, B, C, D, E, or F) are significantly different ( P
Article Snippet: Cells were plated on poly-D-lysine/laminin (Sigma-Aldrich, St. Louis, MO) coated 24-well plate and cultured in serum-free Dulbecco's Modified Eagle Medium: Nutrient Mixture F-12 (DMEM/F-12) (Invitrogen, Gaithersberg, MD) supplemented with 2% B27, 20 ng/ml epidermal growth factor, and 20 ng/ml basic fibroblast growth factor along with antibiotics.
Techniques: In Vitro, Transduction, Cell Culture, Staining