Transdifferentiated Mesenchymal Stem Cells
|Species||Human (H. sapiens)|
|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.|
1. For for-profit organizations and corporations, please contact [email protected] for pricing of this item.
2. Sale of this item is subjected to the completion of a Material Transfer Agreement (MTA) by the purchasing individual/institution for each order. If you have any questions regarding this, please contact us at [email protected].
3. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers or ordering RNA (0.5 μg, Cat.# C207, $450.00) or cell lysate (100 μg, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final.
4. We recommend live cell shipments for ease of cell transfer and this option can be requested at the time of ordering. Please note that the end-user will need to evaluate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical location. In addition, we thoroughly test our cell lines for freeze-thaw recovery. If frozen cells were received and not recovered in your lab under the exact, specified conditions (using recommended culture vessel, media, additional supplements, and atmospheric conditions), a live cell replacement is possible at a cost (plus shipping).
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|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.
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