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| Status |
Public on Aug 31, 2015 |
| Title |
Impacts of reprogramming method and culture conditions in trophoblast differentiation of induced pluripotent stem cells |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
Trophoblast differentiation from human ESC has been achieved by exposing the cells to BMP4 with or without supplementation of ALK4/5/7 inhibitor (A83-01) and FGF2 signaling inhibitor (PD173074) (BAP). Here the two differentiation conditions, BMP4 and BAP were applied to two sets of human PSC lines, H1 ESC and iPSC that latter was generated by DOX-inducible lentiviral (V) transductions of umbilical cord mesenchymal cells. The V-iPSC showed residual transgene expressions from the viral vectors in DOX-free culture condition. When the both ESC and iPSC lines were differentiated simultaneously, similar time dependent morphological changes were observed but BMP4 treated V-iPSC showed a minor yet consistent lag in the differentiation progression compared to BMP4 treated hESC.
Although both differentiated ESC and V-iPSC showed dominant trophoblast phenotypes, the BMP4 treated V-iPSC also expressed gene markers consistent with the presence of mesoendoderm. The BAP condition provided more efficient differentiation than BMP4 alone, and the BAP-differentiated iPSC and ESC never expressed mesoendoderm markers.
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| Overall design |
Five samples, one control of undifferentiated V-iPSC (FGF2) and four differentiated samples included two H1 ESC (treated with BMP4 or BMP4+A83-01) and similarly treated two V-iPSC were analyzed. Trophoblast differentiation was conducted with BMP4 (10 ng/ml) with or without supplementation of ALK4/5/7 inhibitor (A83-01; 1 μM) to H1 ESC and V-iPSC, respective reagents were added to FGF2-free MEF-CM from the second day culture of following passages for up to six additional days.
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| Contributor(s) |
Ezashi T, Roberts RM |
| Citation missing |
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| Submission date |
Dec 17, 2014 |
| Last update date |
Jan 09, 2018 |
| Contact name |
Toshihiko Ezashi |
| E-mail(s) |
ezashit@missouri.edu
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| Phone |
573 884-9601
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| Organization name |
University of Missouri-Columbia
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| Department |
Animal Sciences
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| Lab |
R.M. Roberts
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| Street address |
240a CS Bond Life Sciences Center
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| City |
Columbia |
| State/province |
MO |
| ZIP/Postal code |
65211 |
| Country |
USA |
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| Platforms (2) |
| GPL10332 |
Agilent-026652 Whole Human Genome Microarray 4x44K v2 (Feature Number version) |
| GPL13497 |
Agilent-026652 Whole Human Genome Microarray 4x44K v2 (Probe Name version) |
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| Samples (20)
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| Relations |
| BioProject |
PRJNA270612 |
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