Specifications
| SKU | T2027 |
|---|---|
| Species | Mouse (M. musculus) |
| Species description | PND 35 Oct4-GFP Mice |
| Tissue/Organ/Organ System | Reproductive |
| Pluripotency | Positive for markers for alkaline phosphatase, and SSEA-1 by immunocytochemistry. It is capable of differentiating to cardiomyocytes or glial cells. |
| Caution | For Research Use Only, not for therapeutic or diagnostic purposes. |
| Cell Type | Ready-to-Use Stem Cells |
| Propagation Requirements | Cells are grown on Mouse Embryonic Fibroblast Feeder Cells (Cat. No. T2001). The basal medium for this cell line is Prigrow III medium available at abm, Cat. No. TM003. To make the completed growth medium, add the following components to the base medium to the stated final concentration: 15% fetal bovine serum (TM999), MEM nonessential amino acids (ThermoFisher; 11140-050), 2 mM L-glutamine (G275), 50 µM β-mercaptoethanol (CH045), 10 3 U/ml ESGRO, and Penicillin/Streptomycin (G255). Carbon dioxide (CO2): 5%, Temperature:37.0°C.. For in vitro glial differentiation: gPS cells aggregated to embryoid bodies. The base medium for this cell line is Prigrow III medium available from ABM (TM003). To make the completed growth medium, add the following components to the base medium to the stated final concentration: 15% fetal bovine serum (TM999), MEM nonessential amino acids (ThermoFisher; 11140-050), 2 mM L-glutamine (G275), 50 µM β-mercaptoethanol (CH045), 10 3 U/ml ESGRO (EMD Millipore; ESG1107), and Penicillin/Streptomycin (G255). Atmosphere: air: 95%, CO₂: 5%; Temperature: 37.0°C. Culture the cells with the complete growth media in presence of 10 ng/ml FGF2 (Z101455) and 20 ng/ml EGF (Z100135), followed by propagating cells in complete growth media with 10 ng/ml FGF2 (Z101455) and 10 ng/ml PDGF (Z100365) for 4 days. Differentiation is induced via 4-day growth factor withdrawal in presence of 30 ng/µl 3,3,5-tri-iodothyronine (T3; Sigma) and 200 µM ascorbic acid (AA; Sigma) in complete growth media. Atmosphere: air: 95%, CO₂: 5%; Temperature: 37.0°C.For in vitro cardiomyocytes differentiation: gPS cells cultivated in hanging drops as aggregates to embryoid bodies for 48 hours. The base medium for this cell line is Prigrow III medium available from ABM (TM003). To make the completed growth medium, add the following components to the base medium to the stated final concentration: 20% fetal bovine serum (TM999), MEM nonessential amino acids (ThermoFisher; 11140-050), 2 mM L-glutamine (G275), 50 µM β-mercaptoethanol (CH045), 10 3 U/ml ESGRO (EMD Millipore; ESG1107), and Penicillin/Streptomycin (G255). Atmosphere: air: 95%, CO₂: 5%; Temperature: 37.0°C. After 48 hours, resuspend cells with 5 ml complete growth media and propagate cells as suspension in bacteriological petri dish for 5 days. Then plate embryoid bodies separately onto gelatin-coated 24 wells. Once attached, cardiomyocytes should appear between epithelial-like embryoid body cell outgrowth. |
| Unit quantity | Vial |
| Disclaimer |
1. For for-profit organizations and corporations, please contact quotes@abmgood.com 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 licensing@abmgood.com. 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). 5. All of abm‘s cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic application(s). Please contact a technical service representative for more information. 6. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature and provided for informational purposes only. abm does not warrant that such information has been shown to be accurate. 7. abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the “Warranty Period.” |
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 ~3×10^5 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 3×10^5. Afterwards, please plate the entire IPSC into one well at 10^6 concentration.
4
- Igelmund, P et al. “Action potential propagation failures in long-term recordings from embryonic stem cell-derived cardiomyocytes in tissue cultur” Pflugers Arch 437(5):669-79 (1999). PubMed: 10087143.
- Brustle, O et al. “Embryonic stem cell-derived glial precursors: a source of myelinating transplants” Science 285(5428):754-6 (1999). PubMed: 10427001.
- Ko, K et al. “Induction of pluripotency in adult unipotent germline stem cells” Cell Stem Cell 5(1):87-96 (2009). DOI: 10.1016.
- Maltsev, VA et al. “Embryonic stem cells differentiate in vitro into cardiomyocytes representing sinusnodal, atrial and ventricular cell types” Mech Dev 44(1):41-50 (1993). PubMed: 8155574.
