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Print Version |
| Safety |
BioSafety Level II |
| Organism |
Human |
| Source Organ |
Fetal Lung Fibroblasts (MRC-5) |
| Picture |
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| Image |
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| Concentration |
106 Cells/ml |
| Induction |
iPS-14 was generated using lentiviral transduction of human fetal lung fibroblasts with cMYC, KLF4, OCT4 and SOX2. |
| Pluripotency |
Positive for markers Nanog, Tra-1-60, Tra-1-80 and SSEA-3 by immunofluorescence and flow cytometry. Capable of differentiating into derivatives of the 3 germ layers. |
| Propagation |
The base medium for this cell line is Prigrow IV medium available from ABM (TM004). To make the complete growth medium, add the following components to the base medium: serum replacer to a final concentration of 20%; b-FGF to a final concentration of 6ng/ml; 1 mM L-glutamine and 2-mercaptoethanol solution; 1X Non-essential amino acids. Sterilize the media through a 0.22 micron filter before use. This cell line needs to be cultured on mouse embryonic fibroblasts (ABM T2001) feeder cells. Atmosphere:air: 95%, CO2: 5%; Temperature: 37.0°C. |
| Subculturing |
1. Remove and discard culture medium.
2. Add 2.0mL of Dispase solution to flask and incubate the cells at 37°C to facilitate dispersal. Observe cells under an inverted microscope until cell layer is dispersed.
3.Transfer the dispase solutions containing colonies into a conical tube.
4. Centrifuge cells at 1000rpm for 3 minutes to pellet.
5. Gently aspirate out the solution, leaving pellet undisturbed.
6. Resuspend pellet in fresh culture medium and plate on MEF feeder plates.
7. Incubate cultures at 37°C. |
| Storage |
-180°C |
| Shipping |
On Dry Ice |
| Notes |
For Research Use Only, not for therapeutic or diagnostic purposes. |
| References |
1. Stuart M Chambers, Christopher A Fasano, Eirni P Papapetrou, Mark Tomishima, Michel Sadelain and Lorenz Studer. Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling. Nat Biotechnol. 26, 275-280 (2009).
2. Papapetrou EP, Tomishima MJ, Chambers SM, Mica Y, Reed E, Menon J, Tabar V, Mo Q, Studer L, Sadelain M. Stoichiometric and temporal requirements of Oct4, Sox2, Klf4, and c-Myc expression for efficient human iPSC induction and differentiation. Proc Natl Acad Sci U S A. Aug 4;106(31):12759-64. (2009). |
