High-throughput screening for bioactive molecules using primary cell culture of transgenic zebrafish embryos
- Authors
- Huang, H., Lindgren, A., Wu, X., Liu, N.A., and Lin, S.
- ID
- ZDB-PUB-121004-13
- Date
- 2012
- Source
- Cell Reports 2(3): 695-704 (Journal)
- Registered Authors
- Huang, Haigen, Lin, Shuo, Liu, Ning-Ai
- Keywords
- none
- MeSH Terms
-
- Animals
- Animals, Genetically Modified/genetics
- Animals, Genetically Modified/metabolism*
- Cells, Cultured
- Drug Evaluation, Preclinical/methods*
- Embryo, Mammalian/cytology
- Embryo, Mammalian/metabolism
- Embryo, Nonmammalian/cytology*
- Embryo, Nonmammalian/metabolism*
- Embryonic Stem Cells/cytology
- Embryonic Stem Cells/metabolism
- Gene Expression Regulation/drug effects
- Gene Expression Regulation/physiology
- Green Fluorescent Proteins/biosynthesis
- Green Fluorescent Proteins/genetics
- Human Umbilical Vein Endothelial Cells/cytology
- Human Umbilical Vein Endothelial Cells/metabolism
- Humans
- Mice
- Organ Specificity
- Zebrafish/genetics
- Zebrafish/metabolism*
- PubMed
- 22999940 Full text @ Cell Rep.
Transgenic zebrafish embryos expressing tissue-specific green fluorescent protein (GFP) can provide an unlimited supply of primary embryonic cells. Agents that promote the differentiation of these cells may be beneficial for therapeutics. We report a high-throughput approach for screening small molecules that regulate cell differentiation using lineage-specific GFP transgenic zebrafish embryonic cells. After validating several known regulators of the differentiation of endothelial and other cell types, we performed a screen for proangiogenic molecules using undifferentiated primary cells from flk1-GFP transgenic zebrafish embryos. Cells were grown in 384-well plates with 12,128 individual small molecules, and GFP expression was analyzed by means of an automated imaging system, which allowed us to screen thousands of compounds weekly. As a result, 23 molecules were confirmed to enhance angiogenesis, and 11 of them were validated to promote the proliferation of mammalian human umbilical vascular endothelial cells and induce Flk1+ cells from murine embryonic stem cells. We demonstrated the general applicability of this strategy by analyzing additional cell lineages using zebrafish expressing GFP in pancreatic, cardiac, and dopaminergic cells.