Abstract
Little is known about genes that induce stem cells differentiation into astrocytes. We previously described that heat shock protein 27 (HSP27) downregulation is directly related to neural differentiation under chemical induction in placenta-derived multipotent stem cells (PDMCs). Using this neural differentiation cell model, we cross-compared transcriptomic and proteomic data and selected 26 candidate genes with the same expression trends in both omics analyses. Those genes were further compared with a transcriptomic database derived from Alzheimer’s disease (AD). Eighteen out of 26 candidates showed opposite expression trends between our data and the AD database. The mRNA and protein expression levels of those candidates showed downregulation of HSP27, S100 calcium-binding protein A16 (S100A16) and two other genes in our neural differentiation cell model. Silencing these four genes with various combinations showed that co-silencing HSP27 and S100A16 has stronger effects than other combinations for astrocyte differentiation. The induced astrocyte showed typical astrocytic star-shape and developed with ramified, stringy and filamentous processes as well as differentiated endfoot structures. Also, some of them connected with each other and formed continuous network. Immunofluorescence quantification of various neural markers indicated that HSP27 and S100A16 downregulation mainly drive PDMCs differentiation into astrocytes. Immunofluorescence and confocal microscopic images showed the classical star-like shape morphology and co-expression of crucial astrocyte markers in induced astrocytes, while electrophysiology and Ca2+ influx examination further confirmed their functional characteristics. In conclusion, co-silencing of S100A16 and HSP27 without chemical induction leads to PDMCs differentiation into functional astrocytes.Graphical abstract
Highlights
Due to their properties of self-renewal and differentiation, stem cells, including mesenchymal stem cells and induced pluripotent stem cells, are promising for regenerative medicine [1]
The results indicate that silencing of heat shock protein 27 (HSP27) and S100 calcium-binding protein A16 (S100A16) leads to placenta-derived multipotent stem cells (PDMCs) differentiation into astrocytes
We previously demonstrated that HSP27 downregulation enhances PDMC differentiation into glutamatergic neurons under chemical induction [4]
Summary
Due to their properties of self-renewal and differentiation, stem cells, including mesenchymal stem cells and induced pluripotent stem cells (iPSCs), are promising for regenerative medicine [1]. In order to investigate the crucial genes with the same expression trends, we set the exclusion criteria as 1.28 in log notation Using this strategy, we narrowed the list of gene candidates to nine upregulated genes and 17 downregulated genes at both the mRNA and protein levels. (C and D) mRNA expression of the selected genes from the double-cross-comparison strategy verified by qPCR in the PDMCs induced neural cell model (C for upregulation and D for downregulation). From those results, PDGFRA, S100A16, PLCB3, HSP27, and MT1E showed the same trends for protein expression as for mRNA expression; we kept these genes in the candidate list.
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