You are not logged in.
now: Wed Oct 18 21:35:53 2017 ... mod: Fri Mar 21 06:45:49 2014
Log in
CultureMedium
CultureMedium

Mammalian Tissue Culture Cell Growth Medium

There are a tremendous variety of recipies for growth medium for tissue culture cells. Some cell lines require that specific growth factors, such as insulin, be added to promote either their growth or phenotypic maintenence, however, most cell lines will either grow well, or can be adapted to grow well, in a fairly generic type of medium with 10% final concentration of fetal bovine serum. Some recipes follow:

final FBS
10%
15%
20%

10%
15%
DME-HG
500 ml
500 ml
500 ml

1000 ml
1000 ml
DMEM media with high glucose
FBS
57.5 ml
91.5 ml
130 ml

115 ml
183 ml
fetal bovine serum -- contains growth factors -- do NOT heat inactivate
NEAA
5.75 ml
6.1 ml
6.5 ml

11.5 ml
12.2 ml
non-essential amino acids from 100x stock
P/S
5.75 ml
6.1 ml
6.5 ml

11.5 ml
12.2 ml
pennicilin/streptomycin from 100x stock -- prevents bacterial contamination
l-Gln
5.75 ml
6.1 ml
6.5 ml

11.5 ml
12.2 ml
l-glutamine from 100x stock
plasmocin
57.5 μl
61 μl
65 μl

115 μl
122 μl
prevents mycoplasma contamination (stock 25 mg/ml)

Coriell's Mix: Coriell says to not add extra amino acids or vitamins1
final FBS
5%
10%
15%
MEM (eagle/earle)
500 ml
500 ml
500 ml
minimal essential medium with earle's salts not modified minimal essential medium
FBS
26.9 ml
56.8 ml
90 ml
fetal bovine serum, NOT heat inactivated
P/S
5.4 ml
5.7 ml
6 ml
pennicillin/streptomycin, 100x
l-Gln
5.4 ml
5.7 ml
6 ml
l-glutamine, 100x
plasmocin
53.8 μl
56.8 μl
60.2 μl
anti-mycotic

Shake well to mix and store at 4C.

Fetal bovine serum, l-glutamine, and penn/strep stocks should be kept frozen for long-term storage at -20C. Working bottles of these solutions can be acceptably stored refrigerated at 4C. The stock bottles of DMEM and non-essential amino acids can be stored long-term at 4C.

Converting 10% FBS medium to 15% FBS medium
Add 1/17th volume of FBS to medium already prepared at 10% FBS to bring the final FBS concentration to 15%. Eg: Add 588 μl FBS to 10 mL of 10% FBS medium

pH indicators
Very often, media comes with the pH indicator phenol red pre-added. phenol red is:

Growing cultures are healthy at around pH 7.4. As nutrients and the buffering capacity of the medium becomes exhaused through consumption and the build-up of metabolic waste products, the pH of the culture will gradually drop. The medium should be changed and/or the culture split at orange-red or lower pH colors. The 5% CO2 typcially present in tissue culture incubators maintains pH stability through a carbonate based buffering system:
H2O + CO2 ⇌ H2CO3 ⇌ H+ + HCO3- ⇌ 2H+ + CO3-2

1 Really. I talked to them directly and despite confusing and contradictory instructions on their web site, this is what they actually say to do.

Comments


I use the Leptin green coffee 1000 and it has hepled me lose weight, I no longer use my asthma medications, I no longer use cholesterol meds, I no longer use my hypertension medication. I am allergic to caffeine in the regular coffee and I am not allergic to the green coffee because green coffee is the young coffee beans they have used. I am also a heart patient and my heart does not race with the Leptin Green Coffee 1000
edit this text