Media conditioning for improving gene delivery efficiency to differentiating embryonic stem cells
The present invention provides systems and methods for improving the efficiency of a transient gene delivery system to differentiating embryonic stem (ES) cells by serum starving the targeted cells for one to three days prior to transfection. Such a serum starvation surprisingly resulted in increased expression of a constitutively-controlled plasmid from 50.4% to 83.2% of the population and increased expression of a promoter/enhancer controlled plasmid from ˜1.4% to ˜3.7% of the population.
1. A method for delivering a non-viral plasmid vector to a differentiating embryonic stem cell comprising:
culturing a differentiating human or murine embryonic stem cell in a serum starved condition for about 1 to about 3 days,
delivering to the cell cultured in the serum starved condition a non-viral plasmid vector encoding and capable of expressing a bioactive agent; and
expressing the bioactive agent within the differentiating embryonic stem cell.
2. The method of claim 1 wherein the cell is a differentiating human embryonic stem cell.
3. The method of claim 2 further comprising the step of isolating the embryonic stem cell from an embryoid body prior to the culturing step.
4. The method of claim 1 wherein the cells are cultured on IMDM media having 0.5% or less of fetal bovine serum.
5. The method of claim 1 wherein the cell is a differentiating murine embryonic stem cell.