Figure 3

Gene expression profiles of hMSC-spheroids grown in dynamic 3D-culture. (a) A cluster gram of the PCR array displayed the expression profile of 84 key genes related to the nature of hMSCs, which relatively differed between the 2D- and 3D-MSCs. The gene expression profile of the 3D-MSCs on D1 appeared to be transitional, as they were in the process of forming 3D-spheroids during the 7-day culture period. (b) Comparison of gene expression related to stemness and MSC markers. FGF2, LIF, and POU5F1 were expressed in all groups, while FGF2 and LIF generally decreased upon formation of hMSC-spheroids and subsequently increased over culture time. A variety of hMSC marker genes was highly expressed in 3D-MSCs, showing levels and patterns comparable to those in 2D-MSCs. (c–e) Gene expression representing hMSC’s attributes with average Ct values below 30 are presented by scatter plots and compared between groups. Upon formation of hMSC-spheroids (D1), GDF15 and TGFB3 were upregulated by approximately 40-fold compared to the 2D control, whereas BMP4 was downregulated by approximately 60-fold (c). As our dynamic 3D hMSC-culture progressed up to D7, IL1B, BDNF, and BMP2 were upregulated by more than 30-fold while COL1A1 was downregulated by approximately 50-fold, compared to the early stage of 3D-MSCs on D1 (d). Comparison between 3D-MSC on D7 and 2D-MSCs showed that IL1B and GDF15 were upregulated by approximately 40- and 90-fold, respectively. Particularly, BMP2 was extensively upregulated by approximately 230-fold (e).