Journal: Stem Cells International

Article Title: Efficient Nonviral Transfection of Human Bone Marrow Mesenchymal Stromal Cells Shown Using Placental Growth Factor Overexpression

doi: 10.1155/2018/1310904

Figure Lengend Snippet: Sustained hBM-MSC growth and secretion of placental growth factor after TransIT-2020 transfection. (a) Representative 20X phase contrast images showing cell morphology of hBM-MSCs with reagent only control (R) and the 4h-OM condition with either the pCMV6 empty control or pCMV6-PlGF vector. Scale bars represent 200 μ m. (b) Quantification of PlGF secretion per well in a 6-well plate (pg/ml) in the hBM-MSC cell-conditioned media 24 hours after transfection by ELISA. (c) Amount of viable cells present 24 hours after transfection expressed as a percentage of initial seeding density in a 6-well plate. (d) Percentage of viable cells after transfection in a 6-well plate. (e) Quantification of PlGF secretion (pg/ml) per T75 flask. (f) Amount of viable cells present 24 hours after transfection in a T75. (g) Percentage of viable cells after transfection in a T75. (h) Secretion of PlGF (pg/ml) throughout the 7 days after transfection. (i) Amount of cell growth up to 7 days after transfection, expressed as a percentage of viable cells measured divided by the initial seeding of 37,500 cells. Results from 3 (a–d) or 4 (e–i) independent experiments with technical duplicates. Error bars represent S.E.M. Statistical significance was obtained using a one-tailed t -test for (b) and a two-way ANOVA with a Bonferroni post hoc analysis for (f–g). ∗ p < 0.05 and ∗∗ p < 0.01.

Article Snippet: pCMV6-AC-GFP (cat.#PS100010) and pCMV6-AC-PlGF (cat.#SC320206) plasmids were purchased from OriGene (Rockville, MD, USA). pCMV6-AC-GFP was amplified using Alpha-Gold competent cells (cat.#CC100001, OriGene) and the pCMV6-AC empty vector was generated from glycerol stocks (cat.#PS200020-GLY, OriGene).

Techniques: Transfection, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, One-tailed Test