Commercial dishes with gelatin-free microstructured inserts for elongated stem cell self-renewal and pluripotency

Abstract

Summary: Here, we report the scalable fabrication of 2i-functionalized micro-pyramid-array (μPyA/+2i) inserts for use in commercial multi-well plates, as the alternative cultivation platform for maintaining long-term self-renewal and pluripotency of multiple mESCs and mouse induced pluripotent stem cells. Relevant evidence including cell morphology characterization increased alkaline phosphatase activity, high expression of mESC self-renewal markers, decreased levels of differentiation-associated markers, and high proportion of self-renewal marker cells are provided. Further studies demonstrated that μPyA/+2i could cause a higher cell density in mESC colony, and induce gene expression changes. Subsequent studies showed that μPyA/+2i can influence the cytoskeleton and promote cell adhesion through Cldn-7 upregulation. In summary, these μPyA/+2i inserts offer flexible and gelatin-free micro-envriomnets to maintain long-term self-renewal and pluripotency of mESCs. Enabled by the microstructured inserst, the facile stem cell manipulation and transfer among culture dishes will broaden stem cells both in routine and translational applications.

Keywords

• Stem cells research
• Biomaterials