Preparation manual for DIY DMEM and L15 (basal media) for animal cell culture experiments in kitchens - (Translation from original manual in Japanese) This manual is editable by everybody - join to improve!

1. DIY culture media for kitchen experiments
1

2. Background 1: food safe DIY culture
In past experiments, “Green Dakara” sports drink could be blended with
DMEM up to 60% without losing cell culture efficacy.
https://www.slideshare.net/2co/shojinmeat-project-cellular-agriculture-initiative-nov2018
However, all ingredients must be replaced with commonly available
materials to enable kitchens DIY cell culture experiments.
※Research grade reagents = Not approved as food and not commonly available.
Food or food additives that can be ingested (within limits) are commonly available.
Basal media (DMEM, L15) was prepared entirely from commonly
available materials and a DIY cell culture was demonstrated.

3. Background 2: composition of DMEM and L15
The biggest difference between DMEM and L15 is in the minerals.
Other ingredients were assumed common.
Precisely, DMEM and L15 have different amino acid compositions and DMEM uses glucose while L15 uses galactose
DMEM⇒https://www.sigmaaldrich.com/life-science/cell-culture/learning-center/media-formulations/dme.html
L15⇒ https://www.sigmaaldrich.com/life-science/cell-culture/learning-center/media-formulations/l-15.html
DMEM
L15
20 amino
acids
20 amino
acids
Water-
soluble
vitamin B’s
Water-
soluble
vitamin B’s
Glucose
(dextrose)
Glucose
(dextrose)
CaCl2, MgSO4, KCl, NaCl,
NaHCO3, NaH2PO4, Iron
nitrate
CaCl2, MgCl2, MgSO4, KCl,
NaCl, KH2PO4, Na2HPO4

4. Ingredient 1 (Sugar, animo acids, vitamin B)
0.32g per tablet
0.22g amino acid
(+sugar & fiber)
1 tablet (0.2g,
incl. shellac
shell) for 2L.
Glucose for
baking -
Use 4.5g/L
https://item.rakuten.co.jp/fancl-shop/5272/?i
asid=07rpp_10095___e4-jpeuuk00-aev-615
07def-77bf-41df-8809-18631a826757
https://item.rakuten.co.jp/
kenkocom/x252760h/
https://item.rakuten.co.jp/a
uc-garlic/budoutou-3000/

5. Ingredient 2 (Minerals for DMEM)
https://item.rakuten.co.jp/m
amoru/4571474270191/
https://item.rakuten.co.jp/m
atsubayakuhin/10000000/
CaCl2 (calcium
chloride)
NaCl(salt)
https://item.rakuten.co.jp/
jyugo/t-4987306019126/
Half Salt
NaCl/KCl mixed
at 1:1
MgSO4
(Magnesium sulfate)
Mixing at 1:1 molar
quantity makes NaH2PO4
※Iron nitrate not included
・Only miniscule amount is needed.
・Iron nitrate is not commonly available - not a food additive.
・Replaceable with iron (III) chloride?
baking
soda
https://www.amazon.co.jp/%E
3%82%AA%E3%83%AB%E3
%83%88%E3%83%AA%E3%
83%B3%E9%85%B8-%E3%8
3%AA%E3%83%B3%E9%85
%B8%EF%BC%98%EF%BC
%95%EF%BC%85%E3%80%
8C%E9%A3%9F%E5%93%81
%E6%B7%BB%E5%8A%A0%
E7%89%A9%E3%80%8D%E
F%BC%95%EF%BC%90%EF
%BC%90%EF%BD%87/dp/B0
062HL4PU
phosphoric
acid

6. Ingredient 2 (Minerals for L15)
https://item.rakuten.co.jp/
mamoru/4571474270177/
MgCl2
(magnesium
chloride)
https://item.rakuten.co.jp/
kurimoto/100001011/
NaCl(salt)
https://item.rakuten.co.jp/
jyugo/t-4987306019126/
Half salt
1:1 NaCl/KCl
mixture https://www.amazon.co.jp/%E
3%82%AA%E3%83%AB%E3
%83%88%E3%83%AA%E3%
83%B3%E9%85%B8-%E3%8
3%AA%E3%83%B3%E9%85
%B8%EF%BC%98%EF%BC
%95%EF%BC%85%E3%80%
8C%E9%A3%9F%E5%93%81
%E6%B7%BB%E5%8A%A0%
E7%89%A9%E3%80%8D%E
F%BC%95%EF%BC%90%EF
%BC%90%EF%BD%87/dp/B0
062HL4PU
Mixing at 1:1 molar
quantity makes
NaH2PO4
https://item.rakuten.co.jp/m
amoru/4571474270191/
MgSO4
(Magnesium sulfate)
potassium
carbonate
Mixing at 1:1 molar
quantity makes
KH2PO4
baking
soda
phosphoric
acid

7. 1X Solution 20X Solution
Synthesis method
DMEM:
Table salt 0.58g
baking soda 0.37g
Half salt 0.13g
L15:
Table NaCl 0.95g
Half salt 0.10g
125ml water
1 amino acid
tablet
+
99ml water
1 vitamin B pill
General strategy for preparation
・Prepare 1X concentrated solution for salts, glucose, and amino acid, for which large quantity is needed.
・Prepare 20X solution for vitamin B’s and other minerals, for which only trace amount is needed.
・Prepare phosphates from phosphoric acid and mineral salts.
DMEM:
CaCl2 0.50g
MgSO4 0.25g
baking soda
0.19g
Phosphoric acid
diluted 10X 1.4ml
L15:
CaCl2 0.28g
MgSO4 0.20g
MgCl2 0.19g
K2CO3 0.06g
Phosphoric acid
diluted 10X 1.4ml
Mix 1X solution and 20x solution by 19:1, add glucose at 4.5g/L and bon apetit

8. Scenes from the experiment
・Filtration is needed, as the tablet excipients need to be removed.
・Wait until the suspension settles to avoid clogging the filter paper.
Holding position
with a peeler
Filtered 1X
solution

9. Cell culture demonstration
Primary culture of chicken foetal
heart cells by DIY-DMEM:
Successful cell adhesion and
proliferation
(culturing experiment by @thesow41 on twitter)
https://www.nicovideo.jp/watch/sm33104702
※DIY-L15 to be tested

10. Participate in improving this manual!
https://docs.google.com/presentation/d/1cNedlmZcW7JOKB-gM
itL3aIix1YXd5gi0hhlYz1mVqQ/edit#slide=id.g5683e89615_0_0
Source file openly
accessible and editable