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abm Cell Biology Stem Cells and iPSCs Feeder Cells MITC Inactivated Rat Embryonic Fibroblast (REF) Feeder Cells 2 x
MITC Inactivated Rat Embryonic Fibroblast (REF) Feeder Cells 2 x
| CAT.NO | UNIT |
|---|---|
| T2017 | 1×106 cells/ml |
| SKU | T2017 |
|---|---|
| Species | Rat (R. norvegicus) |
| Species description | Rat |
| Tissue/Organ/Organ System | Reproductive |
| Cell Morphology | Fibroblast-like |
| 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 |
| Growth Properties | Adherent |
| 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 | 1x106 cells/ml |
| Storage Condition | -180°C |
| Shipping Conditions | On Dry Ice |
| Caution | For Research Use Only. |
Supporting Protocol
MSDS
QC
Other
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.
- Linta, L et al. "Rat embryonic fibroblasts improve reprogramming of human keratinocytes into induced pluripotent stem cells" Stem Cells Dev 21(6):965-976 (2012). DOI: 10.1089/scd.2011.0026.
- Su, RJ et al. "Efficient Generation of Integration-Free iPS Cells from Human Adult Peripheral Blood using BCL-XL Together with Yamanaka Factors" PLoS ONE 8(5):e64496 (2013). DOI: 10.1371/journal.pone.0064496.
- Su, RJ et al. "Few Single Nucleotide Variations in Exomes of Human Cord Blood Induced Pluripotent Stem Cells" PloS one 4:e59908 (2013). DOI: 10.1371/journal.pone.0059908 . Application: Generation of iPSC.
- Baylink, D.J. et al. "Vectors and methods for the efficient generation of integration/transgene-free induced pluripotent stem cells from peripheral blood cells" Patent : (2014). PubMed: US20140134143.
