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| Status |
Public on Sep 23, 2014 |
| Title |
Small molecules facilitate rapid and synchronous iPSC generation |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
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| Summary |
The reprogramming of somatic cells into induced pluripotent stem cells (iPSCs) upon overexpression of OCT4, KLF4, SOX2 and c-MYC (OKSM) provides a powerful system to interrogate basic mechanisms of cell fate change. However, iPSC formation with standard methods is typically protracted and inefficient, resulting in heterogeneous cell populations. We show that exposure of OKSM-expressing cells to both ascorbic acid and a GSK3-β inhibitor (AGi) facilitates more synchronous and rapid iPSC formation from several mouse cell types. AGi treatment restored the ability of refractory cell populations to yield iPSC colonies, and it attenuated the activation of developmental regulators commonly observed during the reprogramming process. Moreover, AGi supplementation gave rise to chimera-competent iPSCs after as little as 48 h of OKSM expression. Our results offer a simple modification to the reprogramming protocol, facilitating iPSC induction at unparalleled efficiencies and enabling dissection of the underlying mechanisms in more homogeneous cell populations.
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| Overall design |
18 samples were analyzed in total, samples represent bulk cultures of reprogrammable-MEFs (Rep-MEFs) that express OKSM and were supplemented with ascorbic acid and GSK3i during iPS cell induction. Control samples represent similar bulk cultures of reprogrammable-MEFs that express OKSM but were not supplemented with ascorbic acid and GSK3i during iPS cell induction.
GMP-iPSCs were generated using 48 hours of OKSM+AGi induction. Fibroblast-iPSCs were generated using 96 hours of OKSM+AGi induction.
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| Contributor(s) |
Bar-Nur O, Brumbaugh J, Hochedlinger K |
| Citation(s) |
25262205 |
| Submission date |
May 19, 2014 |
| Last update date |
Feb 21, 2018 |
| Contact name |
Ori Bar-Nur |
| E-mail(s) |
ori.bar-nur@mail.huji.ac.il
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| Organization name |
Harvard University
|
| Department |
Center of Regenerative medicine
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| Lab |
Hochedlinger
|
| Street address |
185 Cambridge st.
|
| City |
Boston |
| State/province |
MA |
| ZIP/Postal code |
02114 |
| Country |
USA |
| |
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| Platforms (1) |
| GPL16570 |
[MoGene-2_0-st] Affymetrix Mouse Gene 2.0 ST Array [transcript (gene) version] |
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| Samples (18)
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| GSM1388380 |
GMPs |
| GSM1388381 |
MEFs derived from Reprogrammable-mouse |
| GSM1388382 |
Rep-MEFs express OKSM for 2 days |
| GSM1388383 |
Rep-MEFs express OKSM+ascorbic acid and GSK3i for 2 days |
| GSM1388384 |
Rep-MEFs express OKSM for 4 days |
| GSM1388385 |
Rep-MEFs express OKSM+ascorbic acid and GSK3i for 4 days |
| GSM1388386 |
Rep-MEFs express OKSM for 6 days |
| GSM1388387 |
Rep-MEFs express OKSM+ascorbic acid and GSK3i for 6 days |
| GSM1388388 |
Rep-MEFs express OKSM for 8 days |
| GSM1388389 |
Rep-MEFs express OKSM+ascorbic acid and GSK3i for 8 days |
| GSM1388390 |
Rep-MEFs express OKSM for 10 days |
| GSM1388391 |
Rep-MEFs express OKSM+ascorbic acid and GSK3i for 10 days |
| GSM1388392 |
Rep-MEFs express OKSM for 12 days |
| GSM1388393 |
Rep-MEFs express OKSM+ascorbic acid and GSK3i for 12 days |
| GSM1388394 |
Sorted Thy+ cells from Rep-MEFs expressing OKSM for 3 days |
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| Relations |
| BioProject |
PRJNA248071 |
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