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abm Cell Biology Stem Cells and iPSCs Transdifferentiated Mesenchymal Stem Cells
Transdifferentiated Mesenchymal Stem Cells
| CAT.NO | UNIT |
|---|---|
| T2016 | Vial |
| SKU | T2018 |
|---|---|
| Species | Human (H. sapiens) |
| Tissue/Organ/Organ System | Blood |
| Pluripotency | Positive for markers by immunofluorescence and flow cytometry. tMSC is capable of differentiating into osteocytes, chondrocytes and adipocytes in vitro. tMSC implantation into immunodeficient mice can form bone, tendon, cartilage and skeletal muscles. |
| Caution | For Research Use Only, not for therapeutic or diagnostic purposes. |
| Cell Type | Ready-to-Use Stem Cells |
| Propagation Requirements | The base medium for this cell line is the MSC medium available from ABM (Cat.No. TM022). To make the complete growth medium, add the growth supplements prior to use. Addition of CHIR99021 (Cat. No. G611) to the medium along with culturing the cells on fibronectin-precoated non-TC well plates under hypoxia (3-5%) is recommended to obtain maximal cell proliferation. If the cells are cultured in normoxia (20% O2), the transdifferentiated MSCs may not be able to expand extensively. Temperature: 37.0°C. |
| Unit quantity | Vial |
Supporting Protocol
MSDS
QC
Other
What is the protocol for freezing iPSCs?
1) 4-5 days after cell split, when iPSCs reach a confluency of 30-60%, aspirate out spend medium and add 1ml 0.5mM EDTA/PBS. Incubate cells at 37C for ~5min. 2) When colonies start to detach, gently harvest all the cells into a 5-ml or 15-ml or 50-ml tube by pippeting 1-2 times. Avoid multiple pippetting. Breaking down clumps into single cells may substantially decrease cell survival. May add iPSC medium to harvest all the cells. 3) Spin down at 200g for 2-3 minutes. 4) Carefully aspirate out supernatent and gently resuspend cells in 0.5ml iPSC medium. 5) Add 0.5ml freezing medium (TM023) and ROCK inhibitor. Mix well by flipping the vial and transfer the medium to a cryovial. 6) Finally transfer the vial to -80C freezer for short-term storage (days to weeks). After cells are frozen, you may then transfer the vial to liquid nitrogen tank for long-term storage (years).
What is the recommended seeding density for iPSC?
IPSCs require seeder cells before thawing and plating the IPSC. Seed ~3x10^5 feeder cells in each well of a 6-well plate. After thawing the iPSCs, put all the cells from the vial into 1 well. After several days, you may start splitting the iPSCs into multiple wells depending on how many cells survived. The density mentioned above is for the seeder cells which should be at 3x10^5. Afterwards, please plate the entire IPSC into one well at 10^6 concentration.
