Abstract
BackgroundThree-dimensional (3D) multicellular spheroids of mesenchymal stem cells (MSCs) are generally regarded to have beneficial properties over MSCs in monolayer. Recent literatures have documented that MSCs can self-assemble into 3D spheroids with a greater capacity for differentiation into various cell types when grown on chitosan (CS), a biopolymer. The genomic modulation occurring in these MSC spheroids is thus of essential importance for understanding their uniqueness and therapeutic potentials. In this study, 3D spheroids self-assembled from human umbilical cord MSCs grown on CS membranes were analyzed by mRNA as well as microRNA microarrays, which helped identify the critical signaling events that may alter the cellular functions during the spheroid forming process.ResultsGenes screened from mRNA and microRNA cross-correlation analyses were further confirmed with the quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) analysis. Results revealed the regulation of a significant number of calcium-associated genes, which suggested the crucial role of calcium signaling in CS-derived MSC spheroids. In addition, many genes associated with the multilineage differentiation capacities and those associated with the antiinflammatory and antitumor properties of MSCs were upregulated. The genetic modulation was significantly more remarkable and endured longer for MSC spheroids derived on CS substrates compared to those derived on a non-adherent (polyvinyl alcohol) substrate.ConclusionsBased on the study, the culture substrates used to prepare 3D MSC spheroids may predefine their properties through cell-substrate interaction.Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2164-15-10) contains supplementary material, which is available to authorized users.
Highlights
Three-dimensional (3D) multicellular spheroids of mesenchymal stem cells (MSCs) are generally regarded to have beneficial properties over Mesenchymal stem cells (MSCs) in monolayer
Simultaneous analyses of the messenger RNA (mRNA) and miRNA expression profiles may help narrowing down the signaling events involved in the behavior change of the cells [11]. We examined both mRNA and miRNA expression profiles of the CS substrate-induced 3D spheroids of human MSCs isolated from the umbilical cord, using 2D MSCs on tissue culture polystyrene (TCPS) as a control
Characteristics of human umbilical cord MSCs The expression profile of cell surface markers analyzed by flow cytometry is shown in Additional file 1: Figure S1
Summary
Three-dimensional (3D) multicellular spheroids of mesenchymal stem cells (MSCs) are generally regarded to have beneficial properties over MSCs in monolayer. MSCs attach and spread on CS membranes before they retract their pseudopodia to form multicellular spheroids This self-assembly process is quite different from that occurs in suspension or hanging drop systems, or on nonadherent polymer surfaces. Several genes/proteins have been referred to participate in the process of spheroid formation on CS, including cadherin molecules [8,9], Rho/ Rho-associated kinase (ROCK) [5], and the Wnt molecule [9]. Activations of these proteins were not as evident for spheroids on non-adherent surfaces. A more comprehensive understanding of the genomic profile for CS-derived MSC spheroids is essential for further revealing the substrate-dependent nature of these unique MSC spheroids
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