Our ability to guide differentiation of human pluripotent stem cells (hPSCs) toward desired lineages efficiently and reproducibly in xeno-free conditions is the key to advancing hPSC technology from the laboratory to clinical use. Here we report an engineered biomimetic substrate functionalized with both peptide ligands for α5β1 and α6β1 integrins to support efficient early mesodermal differentiation of human embryonic stem cells (hESCs) when cultured in a differentiation medium containing BMP4. In contrast, mesodermal differentiation is not induced on substrates functionalized with either ligand alone even though the culture medium is identical. Mesodermal differentiation was characterized by immunofluorescent staining, flow cytometric analysis, and RT-PCR analysis of early mesodermal markers Brachyury, Mixl1, and Wnt3. The early mesodermal progenitors derived on the substrate functionalized with both integrin ligands have the normal developmental potential to further differentiate along the hemato-endothelial and cardiac lineages. Immobilized ligands for α5β1 and α6β1 integrins both are permissive, necessary, and sufficient insoluble ligands in this engineered system to support early mesodermal differentiation of hESCs. This synthetic substrate, in conjunction with defined soluble factors, constructs a well-controlled and xeno-free early mesodermal differentiation niche that offers advantages over the previously reported niche constructed with the Matrigel-coated substrate.
|Original language||English (US)|
|Number of pages||9|
|State||Published - Nov 2011|
Bibliographical noteFunding Information:
This research was partially supported by a seed grant from the Institute for Engineering in Medicine of the University of Minnesota. Prof. Kaufman was supported by NIH/NHLBI grants R01 HL077923 and U01 HL100407 .
- Biomimetic material
- Stem cell
- Surface modification