Recipies for Cell Fusion

 

<center> <b><font>Recipies for Cell Fusion</font> </b></center>

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Phosphate-buffered saline (PBS) (0.01 M Na2HPO4 , 0.15 M NaCl, pH 7.4)

 

sodium phosphate dibasic (Na2HPO4)	0.87 g
sodium phosphate monobasic (NaH2PO4 x H2O)	0.54 g
sodium chloride	8.5 g
HPLC water	1 liter

Adjust the pH to 7.4 with 1 N NaOH or 1 M HCl.

Dubelcco Modified Eagle Medium (Complete Media)

 

DMEM (1x liquid tissue culture medium with 4.5 g/l glucose)	70 %	500 ml
NCTC-135 medium	10%	50 ml
Fetal bovine serum	20 %	100 ml
Antibiotic/antimycotic	1%	5 ml
Fungizone	1 %	5 ml

1. Add 50 ml of NCTC-135 medium to a bottle of unopened DMEM media. Mix and transfer 50 ml to a bottle labelled "Washing Media."

2. Add 100 ml of fetal bovine serum, 5 ml antibiotic and 5 ml fungizone to the 500 ml bottle of DMEM media. Mix, label the container "Complete Media", and store in the refrigerator.

Washing Media

 

DMEM media, unopened	500 ml
NCTC-135 media	50 ml

(Note: Washing media is usually aquired by removing media before the addition of fetal calf serum. Fetal calf serum is expensive and necessary for cell growth.)

70% ethyl alcohol

 

95% ethyl alcohol	370 ml
Distilled water	130 ml

Disinfecting solution (for cleaning)

 

Roccall II	5 ml
HPLC water	1 liter

Disinfecting solution (for filling water pan)

 

Roccall II	7.5 ml
HPLC water	4 liter

HAT media

 

HAT supplement, 500 x	0.1 ml
DMEM Complete media	50 ml

(Note: some formulations are 50 x and should be used at the rate of 1 ml / 50 ml media)

HT media

 

HT supplement, 500 x	0.1 ml
DMEM Complete media	50 ml

(NOTE: some formulations are 50 x and should be used at the rate of 1 ml / 50 ml media)