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| Status |
Public on Dec 29, 2014 |
| Title |
Self-renewal of hepatoblasts under chemically defined conditions by reiterative growth factor and chemical screening |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
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| Summary |
Throught an reiterative growth factor and chemical screening, we defined a small molecule and growth factor cocktail, including EGF, glycogen synthase kinase 3 inhibitor (CHIR99021), transforming growth factor β receptor inhibitor (e.g. E-616452), lysophosphatidic acid and sphingosine 1-phosphate, that can sustain long-term self-renewal of murine hepatoblasts under chemically defined conditions. The expandable hepatoblasts (eHBs) by this small molecule and growth factor cocktail expressed a set of genes typical of liver progenitor cells and liver development, and retained the ability to respond to liver developmental cues and produce functional hepatocytes and form bile duct-like structures. Moreover, both early- and late-passage eHBs demonstrated a similar transcriptome profile. Microarray analysis also confirmed that the gene expression of cultured cells resembled hepatoblasts.
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| Overall design |
Expandable hepatoblasts (eHBs) were routinely cultured in basal medium (DMEM/F12 medium supplemented with insulin-transferrin-sodium selenite, 5 mM nicotinamide, 30 μg/ml 2-phospho-L-ascorbic acid and 50 μg/ml bovine serum albumin human recombinant albumin and 1% penicillin/streptomycin including 10 ng/ml EGF, 3 μM CHIR99021, 2 μM E-616452, 5 μM LPA and 0.5 μM S1P, on Matrigel or laminin-coated surface. The culture was split 1:6 using Accutase. Total RNA was prepared from passage 5 and passage 20 of eHBs (passage 5 and passage 20 of eHBs were derived from two independent experiments) using the RNeasy Plus Mini Kit.
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| Contributor(s) |
Li W |
| Citation missing |
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| Submission date |
Sep 04, 2014 |
| Last update date |
Jan 12, 2017 |
| Contact name |
Wenlin Li |
| E-mail(s) |
liwenlin@smmu.edu.cn
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| Organization name |
Second Military Medical University
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| Street address |
800 Xiangyin Road
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| City |
Shanghai |
| ZIP/Postal code |
200433 |
| Country |
China |
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| Platforms (1) |
| GPL7202 |
Agilent-014868 Whole Mouse Genome Microarray 4x44K G4122F (Probe Name version) |
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| Samples (2) |
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| Relations |
| BioProject |
PRJNA260284 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE61119_RAW.tar |
18.7 Mb |
(http)(custom) |
TAR (of TXT) |
| Processed data included within Sample table |
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