Question

9. A post doctoral fellow asks you to prepare a 6 well plate containing 10^5 RAW 264.7 cells/well in OptiMEM media. Which additional reagents need to added to 500 ml OptiMEM media for it to be complete? What is the purpose of using OptiMEM? Describe a scenario in which it is useful to study cells in G0 phase.

Answer 1

Opti-MEM allows one to keep cells in reduced serum conditions during transfection as it contains insulin, transferrin, hypoxanthine, thymidine, and trace elements which make up for lesser serum. Also Opti MEM is better buffered than DMEM with higher amount of HEPES and sodium bicarbonate and this will keep cells happy during transfection procedures.

Multinucleated muscle cells that do not undergo cytokinesis are also often considered to be in the G0 stage. Due to excellent buffering nature of OptiMEM media it provide excellent environment for the study og Go phase.