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7.. After the cells are visibly
v
detacched, imme
ediately add
d the pre-w
warmed OriCellTM
Mouse MSC Growth Medium (6
6 mL for T7
75 flask, 3 mL
m for T25 flask) to
e the trypsinization.
neutralize
8.. Gently piipette the medium
m
ove
er the cells to dislodge
e and resusspend the cells.
c
Repeat 5-6 times un
ntil all the ccells are dis
ssociated frrom the fla
ask and eve
enly
d into a single cell susspension.
dispersed
9.. Transfer the dissocia
ated cells in
nto a 15 mL
m conical tu
ube.
10
0. Centrifug
ge at 250 x g for 5 min
nutes.
11
1. Carefully aspirate off as much of the supe
ernatant as
s possible.
T
12
2. Add 2 mL
L of OriCellTM
Mouse M
MSC Growth
h Medium to the coniccal tube and
d
gently re
esuspend th
he cells tho roughly.
13
3. Plate the cells into appropriate
ain C57BL//6 Mouse
a
e flasks. OriCellTM Stra
FP can be sp
plit at 1:2 o
or other ap
ppropriate ratios.
r
MSCs/GF
14
4. Add an appropriate amount off medium to
o the cells. Incubate tthe cells att 37°C
5 CO2 hu
umidified in cubator.
inside a 5%
N
Note: Care should be taken
t
to av
void introdu
ucing bubbles during pipetting.
Additional Tips
Tiime to Cha
ange Mediium
It is recomm
mended to change
c
the culture me
edium if the
ere are too many dead
d cells
affter passaging.
It is recomm
mended to change
c
the culture me
edium when
never the m
medium bec
comes
ac
cidic, even if the cells do not rea ch 80-90%
% confluency
y. The pH iindicator in the
cu
ulture medium will app
pear yellow
w when acid
dic.
Tiime to Sub
bculture
W
When OriCelllTM Strain C57BL/6
C
Mo
ouse MSCs//GFP are 80-90% con
nfluent, it is
re
ecommende
ed that the
e cells be su
ubcultured.. Do not lett the cells o
overgrow as
s it will
re
esult in contact inhibition.
OriCellTTM Strain
n C57BL/6 Mous
se MSCs
s DIFFERENTIA
ATION USING
T
TM
OriCell DIFFE
ERENTIA
ATION M
MEDIA
ain C57BL//6 Mouse M SCs/GFP ca
an differenttiate into a variety of cell
OriCellTM Stra
ypes including osteocy
ytes, adipoccytes, and chondrocyt
c
tes.
ty
O
Osteogenic Differen
ntiation
M
Materials Required
R
OriCellTM Mes
senchymal Stem Cell O
Osteogenic
c Differentia
ation Mediu
um (Cat. No
o.
UXMX-9002
21)
GU
IMPI0073A2 M
MUBMX-01101
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