Non-CO₂ Medium

For working with live cells at the microscope, when serum is needed for normal division.

To make	mL F10-Ham’s
start with	mL water
add:	g MEM (Sigma M3024-1L)
add:	g HEPES, then pH to 7.3 (initial pH = ~6.3)
add:	mL Na pyruvate 100 mM (thermo SH30239.01)
add:	mL FBS
add:	mL anti/anti

bring to final volume, filter sterilize in BSC, refrigerate

Ingredient	supplier	cat #	1 L	500 mL	250 mL
MEM			13.4	6.7	3.35	g
HEPES			1.3	0.65	0.325	g
Na pyruvate 100 mM			10	5	2.5	mL
FBS	Krackeler	12103C	100	50	25	mL
anti/anti	Fisher	SV30079.01	10	5	2.5	mL

Mix MEM and HEPES in about 70% of the final volume of dH2O.

Initial pH ~6.3
Adjust pH to 7.3
Add appropriate aliquot of FBS, antibiotic/antimycotic, Na pyruvate
Bring to final volume.
Filter sterilize in the tissue culture hood.
Store in the refrigerator.