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ZIRC
ZFIN ID: ZDB-PUB-121004-13
High-throughput screening for bioactive molecules using primary cell culture of transgenic zebrafish embryos
Huang, H., Lindgren, A., Wu, X., Liu, N.A., and Lin, S.
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.
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ABSTRACT

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.

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