mRNA Transfection of Mouse and Human Neural Stem Cell Cultures
Figure 5
Cyclin D1 enhances mouse neurosphere transfection.
A: Adult mouse neurospheres were seeded as single cells and cultured for 3 or 6 days. Day 4 neurospheres and day 6 neurospheres were dissociated, fixed and stained with PI for cell cycle analysis by flow cytometry (black bars) or were plated onto geltrex and stained for Ki67 (white bars). B: Mouse neurospheres were transfected with 500ng EGFP mRNA on day 4 or day 5 after seeding as single cells. Transfection efficiencies were measured by flow cytometry after 24 hours. C: Day 3 adult mouse neurospheres were transfected with 500ng of CyclinD1 or mRNA lipoplexes (lower plot) or LF2000 alone (upper plot). 24 hours after transfection, cells were dissociated and cell cycle profiles obtained by PI staining and flow cytometry. D: Day 6 mouse neurospheres were transfected with 1 µg of mRNA lipoplexes containing either EGFP mRNA or EGFP mRNA plus Cyclin D1 mRNA (500ng each). Transfection efficiency was measured by flow cytometry 24 hours later.