WNT Inhibition and Increased FGF Signaling Promotes Derivation of Less Heterogeneous Primed Human Embryonic Stem Cells, Compatible with Differentiation
| Autor: | Björn Heindryckx, Filip Van Nieuwerburgh, Petra De Sutter, Margot Van der Jeught, Monika Bialecka, Mina Popovic, Björn Menten, Susana M. Chuva de Sousa Lopes, Jasin Taelman, Sharat Warrier, Kuniya Abe, Dieter Deforce, Laurentijn Tilleman |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Cell type Human Embryonic Stem Cells Biology Fibroblast growth factor Transcriptome Embryo Culture Techniques 03 medical and health sciences 0302 clinical medicine Directed differentiation Humans Benzothiazoles Wnt Signaling Pathway reproductive and urinary physiology Cells Cultured Wnt signaling pathway Cell Differentiation Cell Biology Hematology equipment and supplies Embryo Mammalian Embryonic stem cell Cell biology Up-Regulation Wnt Proteins 030104 developmental biology Blastocyst Epiblast embryonic structures Fibroblast Growth Factor 2 Stem cell 030217 neurology & neurosurgery Developmental Biology Signal Transduction |
| Zdroj: | Stem cells and development. 28(9) |
| ISSN: | 1557-8534 |
| Popis: | Human embryonic stem cells (hESCs) hold great value for future clinical applications. However, standard culture conditions maintain hESCs in a primed state, which bears heterogeneity in pluripotency and a tendency for spontaneous differentiation. To counter these drawbacks, primed hESCs have been converted to a naive state, but this has restricted the efficiency of existing directed differentiation protocols. In mouse, WNT inhibition by inhibitor of WNT production-2, together with a higher dose of fibroblast growth factor 2 (12 ng/mL) in DMEM/F12 basal medium (DhiFI), markedly improved derivation and maintenance of primed mouse epiblast stem cells. In this study, we show that DhiFI conditions similarly improved primed hESC traits, such as conferring a primed transcriptional signature with high levels of pluripotency markers and reduced levels of differentiation markers. When triggered to differentiate to neuronal and cardiac lineages, DhiFI hESCs and isogenic primed hESCs progressed similarly. Moreover, DhiFI conditions supported the derivation of hESC lines from a post-inner cell mass intermediate (PICMI). DhiFI-derived hESCs showed less spontaneous differentiation and expressed significantly lower levels of lineage-specific markers, compared to primed-derived lines from the same PICMI. Overall, DhiFI hESCs retained advantages of both primed and naive pluripotency and may ultimately represent a more favorable starting point for differentiation toward clinically desired cell types. |
| Databáze: | OpenAIRE |
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