Abstract
Human adipose-derived stem cells (hADSCs) are types of mesenchymal stem cells (MSCs) that have been used as tissue engineering models for bone, cartilage, muscle, marrow stroma, tendon, fat and other connective tissues. Tissue regeneration materials composed of hADSCs have the potential to play an important role in reconstituting damaged tissue or diseased mesenchymal tissue. In this study, we assessed and investigated the osteogenesis of hADSCs in both two-dimensional (2D) and three-dimensional (3D) culture conditions. We confirmed that the hADSCs successfully differentiated into bone tissues by ARS staining and quantitative RT–PCR. To gain insight into the detailed biological difference between the two culture conditions, we profiled the overall gene expression by analyzing the whole transcriptome sequencing data using various bioinformatic methods. We profiled the overall gene expression through RNA-Seq and further analyzed this using various bioinformatic methods. During differential gene expression testing, significant differences in the gene expressions between hADSCs cultured in 2D and 3D conditions were observed. The genes related to skeletal development, bone development and bone remodeling processes were overexpressed in the 3D culture condition as compared to the 2D culture condition. In summary, our RNA-Seq-based study proves effective in providing new insights that contribute toward achieving a genome-wide understanding of gene regulation in mesenchymal stem cell osteogenic differentiation and bone tissue regeneration within the 3D culture system.
Highlights
IntroductionStem cells have been used for the regeneration of lost tissues because they can differentiate into any type of adult tissue
Adult stem cells have less ability to differentiate into other tissues as compared to ESCs, they are easier to obtain in greater quantities with fewer ethical and technical complications as compared to embryonic stem cells
We focused on the three clusters with highly expressed genes only in the 3D cluster condition in order to determine how the 3D culture conditions differently affected gene expression and biological functions during osteogenic differentiation
Summary
Stem cells have been used for the regeneration of lost tissues because they can differentiate into any type of adult tissue. The supply of these cells is limited due to the ethical problems involved with the collection of these cells and the technical difficulties involved with harvesting them. This reduces their practical applications for tissue regeneration. Tissue regeneration studies have begun to use adult stem cells instead of ESCs. adult stem cells have less ability to differentiate into other tissues as compared to ESCs, they are easier to obtain in greater quantities with fewer ethical and technical complications as compared to embryonic stem cells
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