Abstract
The potential applications of human pluripotent stem cells, embryonic stem (ES) cells, and induced pluripotent stem (iPS) cells in cell therapy and regenerative medicine have been widely studied. The precise definition of pluripotent stem cell status during culture using biomarkers is essential for basic research and regenerative medicine. Culture conditions, including extracellular matrices, influence the balance between self-renewal and differentiation. Accordingly, to explore biomarkers for defining and monitoring the pluripotent substates during culture, we established different substates in H9 human ES cells by changing the extracellular matrix from vitronectin to Matrigel. The substate was characterised by low and high expression of the pluripotency marker R-10G epitope and the mesenchymal marker vimentin, respectively. Immunohistochemistry, induction of the three germ layers, and exhaustive expression analysis showed that the substate was ectoderm-biased, tended to differentiate into nerves, but retained the potential to differentiate into the three germ layers. Further integrated analyses of mRNA and miRNA microarrays and qPCR analysis showed that nine genes (COL9A2, DGKI, GBX2, KIF26B, MARCH1, PLXNA4, SLC24A4, TLR4, and ZHX3) were upregulated in the ectoderm-biased cells as ectoderm-biased biomarker candidates in pluripotent stem cells. Our findings provide important insights into ectoderm-biased substates of human pluripotent stem cells in the fields of basic research and regenerative medicine.
Highlights
Cell therapies can target various human diseases and regenerative medicine applications through the use of different cell lines, including neural stem cells, mesenchymal stem cells, hematopoietic stem cells, and differentiated cells from embryonic stem (ES) cells and induced pluripotent stem cells[1,2,3]
To establish techniques for quality evaluation of human pluripotent stem cells using biomarkers, we first examined the conditions for pluripotency in H9 human ES cells by changing the extracellular matrix
The epithelial-to-mesenchymal transition is observed during ES/induced pluripotent stem (iPS) cell differentiation[21]
Summary
Cell therapies can target various human diseases and regenerative medicine applications through the use of different cell lines, including neural stem cells, mesenchymal stem cells, hematopoietic stem cells, and differentiated cells from embryonic stem (ES) cells and induced pluripotent stem (iPS) cells[1,2,3]. Both ES cells and iPS cells have broad applications in various disease states[2,3]. We evaluated the expression patterns of various markers, examined the differentiation potential of the cells, and performed integrated analyses of mRNA and miRNA microarrays of the cells and exosomes to explore high-quality and reliable biomarkers. Our findings provided insights into the use of biomarker candidates during the earliest stages of ectodermal differentiation
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