|Species||Rat (R. norvegicus)|
|Applications||For Research Use Only. The inactivated REFs have been tested to be significantly better than inactivated MEF in supporting human iPSC reprogramming and long-term undifferentiated proliferation of human iPS cells and human ground state naive pluripotent stem cells. There is no need to change medium at weekend if human iPS or ES cells are split on Friday and cultured on this REF feeder line in iPSC Culture Medium (TM020).|
|Cell Type||Feeder Cells|
|Propagation Requirements||The base medium for this cell line is Prigrow IV medium available at abm, Cat. No. TM004. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999) to a final concentration of 10%, 0.1 mM Non-Essential Amino Acids, and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%.
Carbon dioxide (CO2): 5%, Temperature: 37.0°C
|Unit quantity||1×106 cells/ml|
|Shipping Conditions||On Dry Ice|
|Caution||For Research Use Only.|
Do you sell REFs that have not been inactivated?
Please see the following web page: T2020
How are these cells generated?
There is no virus infection on the REF feeder cells. REFs were isolated from E15 Sprague Dawley rats and cultured in TM004 with 15% FBS.
Are these cells modified by virus?
Yes, this cell line has been transduced with FGF2 and LIF via lentiviruses.
Can T2017 be further passaged after revived in culture dish?
These cells cannot be further passaged as they are mitomycin C treated which will prevent fibroblast cell growth. Once thawed these cells will attach to the vessel and form a monolayer – or a feeder layer – for iPSC growth.
Is there a a protocol for preparing wells with the REF feeder cells for iPSC plating?
One day before plating iPSCs, thaw 1 vial of REF feeder cells. After washing with 10-20 ml culture medium, re-suspend 2 x 106 cells in 12 ml TM004 with 15% FBS. Add 2 ml into each well of a gelatin-coated 6-well plate.