Download User Manual

x
OriCellTM Mouse
M
Adipo
ose-Derived
d Mesenchy
ymal Stem Cell Growtth Medium (Cat. No.
MUXMD-90
0011)
Tha
awing and
d Establish
hing Mouse
e ADSCs
1.
Pre-warm OriCellTM Mouse
M
ADSC
C Growth Medium
M
to 37°C.
3
2.
M
Add 9 mL of OriCellTM
Mouse AD
DSC Growth
h Medium to a 15 mL conical tub
be.
3.
M
Remove th
he cryovial of OriCellTM
Strain C5
57BL/6 Mouse ADSCs ffrom liquid nitrogen.
4.
Quickly thaw the cryo
ovial in a 3
37°C water bath until the
t
last ice crystal dis
sappears.
o finish the thawing prrocedure wiithin 3 minutes. Be ca
areful not to
o submerge
e
Be sure to
the entire vial. Maxim
mum cell vi ability is de
ependent on the rapid
d and comp
plete
thawing off frozen cells.
Notte: Results
s will be les
ss than optiimal if the cells are th
hawed for m
more than 3 minutes.
5.
As soon as
s the cells are
a comple tely thawed
d, disinfect the outsid e of the via
al with 70%
%
v/v ethano
ol.
6.
Use a pipe
ette to trans
sfer the cellls to the co
onical tube containing
g OriCellTM Mouse
M
ADSC Grow
wth Medium
m inside a b
biosafety cabinet. Be careful nott to introduce any
bubbles du
uring the trransfer proccess.
7.
Rinse the vial with 1 mL of the medium to reduce cell loss. Sub
bsequently transfer
pension into
o the conica
al tube.
this 1 mL of cell susp
8.
x the cell su
uspension b
by slowly pipetting
p
up
p and down . Be carefu
ul not to
Gently mix
introduce any bubble
es.
9.
Centrifuge
e the cell su
uspension a
at 250 x g for
f 5 minutes.
10. Carefully aspirate
a
offf as much o
of the supernatant as possible an
nd add 2-3 mL of fresh
h
OriCellTM Mouse
M
ADSC
C Growth M
Medium (pre-warmed to 37°C).
11. Gently re-suspend th
he cells in O
OriCellTM Mo
ouse ADSC Growth Me
edium.
12. Plate the cells
c
into a T25 flask a
and add a sufficient
s
OriCellTM Mou
use ADSC Growth
G
Medium. Gently
G
rock the culture
e flask to evenly distribute the ce
ells.
13. Incubate at
a 37°C in a 5% CO2 h
humidified incubator.
14. The next day,
d
change
e the mediu
um with fre
esh growth medium (p
pre-warmed
d to 37°C)..
15. Change th
he growth medium
m
eve
ery three days thereaffter.
16. When the cells are ap
pproximate
ely 80-90%
% confluent, they can b
be dissociatted with
0.25%Trypsin-0.04%
%EDTA and passaged.
Notte: Changiing Mediu
um
1.
Warm an appropriate
a
e amount o f medium to
t 37°C in a sterile con
ntainer. Re
eplace the
spent med
dium with the pre-warrmed, fresh
h medium. Once comp
pleted, retu
urn the
flask to the incubator.
2.
eated warm
ming and co
ooling of the medium. If the entiire contentt is not
Avoid repe
needed forr a single procedure,
p
ttransfer on
nly the requ
uired volum
me to a ster
rile
secondary
y container..
IMPI0020A2M
MUBMDͲ01001 Page5of14