Aggregation morphology expression of pluripotency related genes of 3D-spheroid RA-hMSCs. (A)Stereo microscope showing the time course of the aggregation of 25,000 RA-hMSCs into a spheroid in a hanging drop. Time-lapse microscopy demonstrated that RA-hMSCs cultured in hanging drops formed a loose network at 24h and then numerous small aggregate that gradually coalesced into a single central spheroid along the lower surface of the drop (scale bar, 500 μm). (B) The size of single cells derived from re-isolated 2D- and 3D-spheriod culture hMSCs were observed. The cells released from spheroids by trypsinization were nearly half the diameter and approximately one-fourth the volume of RA-hMSCs derived from 2D cultured. Arrows indicate single cells (scale bar, 100 μm). (C) Relative mRNA levels of pluripotent related genes (OCT4, SOX2, and NANOG) by qRT-PCR analysis from 2D cultured and 3D-spheroids cultured hMSCs. All data was normalized against YWHAZ and presented the mean ± SD (***p < 0.001).
pISSN: 2671-4639
