CFU-F assays and differential potential of dental mesenchymal cells of <i>Wnt1-Cre/YFP</i> and <i>Mesp1-Cre/YFP</i> mice.

<p>(A) Protocol of the CFU-F assays and fibroblastic colonies obtained from <i>Wnt1-Cre/YFP</i> mice. Scale bars, 200 µm. (B) Numbers of colonies induced from dental mesenchymal cells including both YFP⁺ and YFP⁻ cells of E13.5 and 2-day-old <i>Wnt1-Cre/YFP</i> and <i>Mesp1-Cre/YFP</i> mice. Values represent the mean (SD) of triplicate cultures of two independent experiments. Y⁺, YFP⁺ colonies; Y⁻, YFP⁻ colonies. (C) Percentages of YFP⁺ and YFP⁻ colonies induced from dental mesenchymal cells of <i>Wnt1-Cre/YFP</i> and <i>Mesp1-Cre/YFP</i> mice. Figures within parentheses indicate the number of YFP⁺ colonies of the total number of colonies. Values represent percentages and colony numbers of triplicate cultures of two independent experiments (<i>n</i> = 6/group). (D) Differentiation potential of NC-derived and mesoderm-derived dental mesenchymal cells into adipocytes, osteoblasts, and chondrocytes. Cells from YFP⁺ or YFP⁻ colonies using dental mesenchymal cells from <i>Wnt1-Cre/YFP</i> or <i>Mesp1-Cre/YFP</i> mice were collected using a cell sorter. YFP expression in each cell preparation is shown in the lower panel. The cultured cells were stained with oil red O (OilR), alizarin red (ALZ), and anti-type II collagen (Col II) antibody to detect adipocytes, osteoblasts, and chondrocytes, respectively. ST2 and ATDC5 cells were the positive controls. The experiments were repeated twice and one representative experiment is presented. Scale bars, 200 µm in OilR and Col II and 1 mm in ALZ.