Fig. 2

Fabrication and characterization of ExpLBM-derived chondrocyte sheets. A A diagram of ExpLBM-derived chondrocyte sheet fabrication. At the end of 2-DCI, ExpLBM-derived chondrocytes were dissociated and seeded onto a temperature-responsive culture insert. B Alcian blue staining after 2-DCI (scale bar: 200 µm). C Fabrication of 414C2 or Ff-KVs09 ExpLBM-derived sheet-like structures. Images were acquired at the specified time points (scale bar: 200 µm). D Quantitative reverse-transcription-polymerase chain reaction (qRT-PCR) analysis of chondrocyte marker genes in ExpLBM-derived chondrocyte sheets. Total RNAs were extracted, and the expression levels of each gene were compared between ExpLBM and chondrocyte sheets. All values were normalized to TBP mRNA levels (n = 3, three biologically independent experiments). E Histological examination of 414C2 or Ff-KVs09 ExpLBM-derived chondrocyte sheets. Chondrocyte sheet sections were stained with hematoxylin and eosin (H&E), Alcian blue, or Safranin O, or with antibodies against SOX9, COL2, ACAN, COL1, and RUNX2 (scale bars: 200 µm)