title: Combined activin A/LiCl/Noggin treatment improves production of mouse embryonic stem cell-derived definitive endoderm cells.
abstract: Induction of definitive endoderm (DE) cells is a prerequisite for the whole process of embryonic stem (ES) cells differentiating into hepatic or pancreatic progenitor cells. We have established an efficient method to induce mouse ES cell-derived DE cells in suspension embryonic body (EB) culture. Similar to previous studies, mouse ES cell-derived DE cells, which were defined as Cxcr4(+) c-Kit(+) , Cxcr4(+) E-cadherin(+) cells or Cxcr4(+) PDGFRa(-) cells, could be induced in the serum-free EBs at Day 4 of induction. The activations of Wnt, Nodal, and FGF signaling pathways in differentiating EBs promoted DE cell differentiation, while activation of BMP4 signaling inhibited the process. In the present study, we found that chemical activation of canonical Wnt signaling pathway by LiCl could synergize with Activin A-mediated Nodal signaling pathway to promote induction of DE cells, and inhibition of Bmp4 signaling by Noggin along with Activin A/LiCl further improved the efficiency of DE cell differentiation. The derived DE cells were proved for their capacities to become hepatic progenitor cells or pancreatic progenitor cells. In conclusion, we significantly improved the efficiency of generating mouse ES cell-derived DE cells by combined Activin A/LiCl/Noggin treatment. Our work will be greatly helpful to generate ES cell-derived hepatic cells and ES cell-derived pancreatic cells for future regenerative medicine.
authors.affiliation: Laboratory of Molecular and Cell Biology, Institute of Biochemistry and Cell Biology, China.
authors.forename: Fuming
authors.initials: F
authors.lastname: Li
authors.forename: Zhiying
authors.initials: Z
authors.lastname: He
authors.forename: Yangfang
authors.initials: Y
authors.lastname: Li
authors.forename: Pingyu
authors.initials: P
authors.lastname: Liu
authors.forename: Fei
authors.initials: F
authors.lastname: Chen
authors.forename: Minjun
authors.initials: M
authors.lastname: Wang
authors.forename: Haiying
authors.initials: H
authors.lastname: Zhu
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Ding
authors.forename: Kirk J
authors.initials: KJ
authors.lastname: Wangensteen
authors.forename: Yiping
authors.initials: Y
authors.lastname: Hu
authors.forename: Xin
authors.initials: X
authors.lastname: Wang
mesh_terms: D028341:Activins / Q000494:pharmacology*; D000818:Animals; D055415:Bone Morphogenetic Protein 4 / Q000378:metabolism; D002352:Carrier Proteins / Q000494:pharmacology*; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D004707:Endoderm / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005434:Flow Cytometry; D007150:Immunohistochemistry; D018021:Lithium Chloride / Q000494:pharmacology*; D008099:Liver / Q000166:cytology / Q000378:metabolism; D051379:Mice; D010179:Pancreas / Q000166:cytology / Q000378:metabolism; D019009:Proto-Oncogene Proteins c-kit / Q000235:genetics; D020796:Receptor, Platelet-Derived Growth Factor alpha / Q000235:genetics / Q000378:metabolism; D019718:Receptors, CXCR4 / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects; D013997:Time Factors; D051153:Wnt Proteins / Q000378:metabolism; D000099176:Noggin Protein
chemical_list: D028341:Activins; D055415:Bone Morphogenetic Protein 4; D002352:Carrier Proteins; D016895:Culture Media, Serum-Free; D018021:Lithium Chloride; D019009:Proto-Oncogene Proteins c-kit; D020796:Receptor, Platelet-Derived Growth Factor alpha; D019718:Receptors, CXCR4; D051153:Wnt Proteins; D000099176:Noggin Protein; C525810:Bmp4 protein, mouse; C514936:CXCR4 protein, mouse; C422405:activin A