Use of Xenofree Matrices and Molecularly-Defined Media to Control Human Embryonic Stem Cell Pluripotency: Effect of Low Physiological TGF-βConcentrations
| Autor: | Isabelle Peiffer, Jacques Hatzfeld, Yi-Ping Zhou, Ma-Lin Li, Marie-Noëlle Monier, Romain Barbet, Antoinette Hatzfeld |
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| Rok vydání: | 2008 |
| Předmět: |
Pluripotent Stem Cells
Left-Right Determination Factors Down-Regulation Biology Mice Transforming Growth Factor beta Albumins Gene expression Animals Humans Gene family Induced pluripotent stem cell Cell Shape Embryonic Stem Cells Cell Proliferation Gene Expression Profiling Mesenchymal stem cell Cell Differentiation Cell Biology Hematology Phenotype Molecular biology Embryonic stem cell Activins Culture Media Extracellular Matrix Up-Regulation Gene expression profiling Chemically defined medium Karyotyping Cytokines Intercellular Signaling Peptides and Proteins Developmental Biology |
| Zdroj: | Stem Cells and Development. 17:519-534 |
| ISSN: | 1557-8534 1547-3287 |
| DOI: | 10.1089/scd.2007.0279 |
| Popis: | To monitor human embryonic stem cell (hESC) self-renewal without differentiation, we used quantitative RT-PCR to study a selection of hESC genes, including markers for self-renewal, commitment/differentiation, and members of the TGF-beta superfamily and DAN gene family. Indeed, low commitment/differentiation gene expression, together with a significant self-renewal gene expres sion, provides a better pluripotency index than self-renewal genes alone. We demonstrate that matrices derived from human mesenchymal stem cells (hMSCs) can advantageously replace murine embryonic fibroblasts (MEF) or hMSC feeders. Moreover, a xenofree molecularly-defined SBX medium, containing a synthetic lipid carrier instead of albumin, can replace SR medium. The number of selected differentiation genes expressed by hESCs in these culture conditions was significantly lower than those expressed on MEF feeders in SR medium. In SBX, the positive effect of a non-physiological concentration of activin A (10-30 ng/mL) to reduce differentiation during self-renewal could also be obtained by physiological concentrations of TGF-beta(100-300 pg/mL). In contrast, these TGF-beta concentrations added to activin favored differentiation as previously observed with TGF-beta concentrations of 1 ng/mL or more. Compared to SR-containing medium, SBX medium promoted down-regulation of CER1 and LEFTIES and up-regulation of GREM1. Thus these genes better control self-renewal and pluripotency and prevent differentiation. A strategy is proposed to analyze, in more physiological, xenofree, molecularly-defined media and matrices, the numerous genes with still unknown functions controlling hESCs or human-induced pluripotent stem cells (iPS). |
| Databáze: | OpenAIRE |
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