TMCiMS

Instruction for usever. 2

Product Description

CiMSTM is a medium for culture of human mesenchymal stem cells (hMSCs). CiMSTM is optimized forthe multiple passage expansion of hMSCs (isolated from Bone Marrow, Adipose Tissue and Dental
Pulp) while still allowing for differentiation into the several desired lineages. Xeno-free or Animal-free
supplement can be selected according to the intended use. In addition, there is no need to pre-coat
culture vessels with any type of attachment matrix before using CiMSTM.
ProductCatalog Number(NIPRO/CSTI)CiMSTM-BM

87-070 / A2G00P05C

Basal media (Animal-free)

500 mL

2-8 ℃ ; Protect from Light

TM-sXF

87-071 / A2G10P1CC

Xeno-free supplement

10 mL-20 ℃ ; Protect from Light*CiMSTM-sAF

87-072 / A2G20P1CC

Animal-free supplement

10 mL-20 ℃ ; Protect from Light

Related ProductCatalog Number(NIPRO/CSTI)

*CiMS

sTI

87-974 / 1210
87-975 / 1220

PBS(-)

87-949, 87-972 / 1102P05, 1102P10

rTE

Components

VolumeComponents

Storage

VolumeRecombinant Trypsin/EDTA solution
Synthetic Trypsin inhibitor

Storage

100 mL2-8 ℃ ; Protect from Light

100 mL

2-8 ℃ ; Protect from Light

500, 1000 mL

1-30 ℃* Please select either Xeno-free or Animal-free supplement.
Xeno-free : It contains human derived component. Any other animal derived component free.
Animal-free : animal derived component free

Storage & thawing of supplement

Preparation of Culture Media

1. CiMSTM instructions: upon arrival, store

1. Decontaminate the external surfaces ofTM

CiMS -BM(basal medium) protected fromlight at 2℃ to 8℃ and CiMSTM-sXF orCiMSTM-sAF at ≤ -20℃ in a freezer.
Quickly thaw a frozen supplement in a
37℃ water bath being careful not tosubmerge the entire bottle. Watch thesupplement closely ; when the wholecontents melt, please take it out
immediately from the water bath. Once
thawed, supplement should be stored
2-8℃ and added to basal medium within
2 weeks. After supplement is added tobasal medium, use immediately.
* Fine particles may be observed but there
is no influence on quality.

the CiMSTM-BM and CiMSTM-sXF or –sAFbottle with 70% v/v ethanol or isopropanol.

2. Add 1/50 volume of CiMSTM-sXF orCiMSTM-sAF to CiMSTM-BM (CiMSTM completemedium).
* Recommend to make necessary volume of themedium just before use.

Initiation of Culture Process
1. Rapidly thaw a frozen cryotube of hMSC in a
37℃ water bath until a small amount of ice
remains.

2. Wipe cryotube with 70% v/v ethanol orisopropanol before opening. Pipet the entire
contents of cryotube into a conical tube.

3. Carefully add 10 mL of pre-warmed (37℃)
CiMSTM complete medium.

4. Centrifuge the tubes at about100 x g for 5
minutes at room temperature and discard thesupernatant.

5. Resuspend the cell pellet in pre-warm (37℃)
CiMSTM complete medium and add the cellsuspension to an appropriate culture vessel(cell culture vessels for adherent cells) at adensity of 0.5 – 1.0 x 104 cells / cm2.

incubator.

the vessel with 0.5 mL of s-TI (Catalog#87-975) at room temperature.

8. Quickly transfer the detached cells to sterile
15 mL centrifuge tube.

9. Rinse the flask with a final 10 mL of

10. Examine the harvested flask underMaintenance
1. Change CiMS

7. After cell released, neutralize the trypsin in

PBS(-)(Catalog #87-949, 87-972) to collectresidual cells, and add this rinse to thecentrifuge tube.

6. Incubate at 37℃, 5% CO2, humidifiedTMthe majority of cells from the culture surface.
If only a few cells detach, you may not have
let them trypsinize long enough.

complete medium every 3days.

microscope to make sure the harvest wassuccessful by looking at the number of cells
left behind. This should be less than 5%.

11. Centrifuge the tubes at about 100 x g for 5

Subculturing
2The following instructions are for a 25 cm flask.
Adjust all volumes accordingly for other sizevessels.
1. Subculture the cells when they are about 85%
confluent.

2. Remove the culture medium from 25 cm2flask.

3. Cover the cells with 0.5 mL rTE (Catalog#87-974) .

4. Place the culture vessel into a 37℃
humidified incubator for 3-5 minutes.
Periodically examine the cell layer
microscopically and check for celldetachment.

5. Allow the trypsinization to continue until

minutes at room temperature and discard
the supernatant.

12. Resuspend the pellet in a minimum volumeof temperature equilibrated CiMSTM
complete medium by gently pipetting up
and down. Count total number of viable
cells.

13. Add the calculated volume of cellsuspension to each flask and gently rock todisperse the cell suspension over the growthsurface.

14. Incubate at 37℃, 5% CO2, humidified
incubator.

Products are for research use only and notintended for human or animal diagnostic ortherapeutic uses unless otherwise stated.

approximately 90% of the cells are roundedup.

6. At this point, tap the flask gently to release

CD19012