Trnascriptome Analysis of 3-D Spheroid Culture of Human Fibroblasts Reveals Potential of the Method in Inducing Stem Cell Reprogramming
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AbstractCompared with 2-D cell culture, 3-D spheroid culture methodology is regarded as more physiologically similar to the native environment in the body. Numerous reports showed spheroid culture of mesenchymal stem cells (MSC) and cancer stem cells acquired enhanced differentiation capacity, sensitivity to chemicals, and elevated expression of differentiation- and stemnessassociated genes. Despite this emerging trend, to date, there is no comprehensive study conducted on the effect of spheroid aggregate cell culture compared to traditional 2-D culture. To investigate into this topic, we used hydrogel-coated culture wells to force human skin fibroblasts forming into spheroid aggregates. Using whole genome RNA sequencing coupled with qPCR validation, we have detected more than 200 differentially expressed genes. Gene Ontology (GO) analysis revealed a majority of the genes are related to extracellular matrix composition and organization, and cell migration. Intriguingly, a number of early to mature stem cell markers and Mesenchymal-to-Epithelial Transition (MET) genes were significantly up-regulated while some of the Epithelial-to-Mesenchymal Transition (EMT) genes were down-regulated, resembling stem cell reprogramming. Similar phenomenon was observed across biological replications, cell types in both human and mouse samples, including somatic cells, MSC and embryonic stem cells. We also found that when used as coating of culture surface, the extracellular matrix of the spheroids could induce expression change of some genes in 2-D cultured fibroblasts in accordance to suspension culture. The result provides an insight in the biological significance of 3-D spheroid culture methodology over the traditional 2-D culture, and the potentials of using spheroid culture in stem cell reprogramming.
All Author(s) ListLO Lok Man, TANG Cheng Cheng, LEE Ka Ho Kenneth
Name of Conference2017 ISSCR Annual Meeting
Start Date of Conference14/06/2017
End Date of Conference17/06/2017
Place of ConferenceBoston Convention and Exhibition Center (BCEC)
Country/Region of ConferenceUnited States of America
LanguagesEnglish-United States

Last updated on 2018-18-01 at 08:32