Multipotent Progenitor Cell Differentiation

Testes were collected from adult C57BL/6 or Tcf21^mCrem:R26R^tdTom mice and dissociated into a single-cell suspension and sorted for SCA1⁺/cKITit⁻ or SCA1⁺/TCF21^lin/cKITit⁻ cells, respectively. For adipogenic differentiation, 3 × 10⁴ cells were plated in a monolayer, cultured for 10 days (Stem ProAdipogenic differentiation kit) and stained with either Oil Red O or Perilipin (1:250, Sigma, Cat#P1873). For chondrogenic differentiation, 5 × 10⁴ cells were plated in micromass, cultured for 21 days (Stem ProChondrogenic differentiation kit) and stained with either Alcian blue or SOX9 (1:250, EMD Millipore, Cat#ABE571). For osteogenic differentiation, 1 × 10⁴ cells were plated in micromass, cultured for 14 days (StemProOsteogenicdifferentiationkit), and stained with either Alizarin red or Osterix (1:250, Abcam, Cat#ab22552)^{96 (link)}.