GSM7916665: MPS, ScRNA-seq; Mus musculus; RNA-Seq
Removing the femoral distal epiphysis and the tibial proximal epiphysis gently, and about 5 mm long the metaphysis region was dissected and crushed with mortar and pestle in FACS buffer (10%FBS in PBS). The isolated metaphysis was cut into small pieces with scissors and was digested with 3mg/ml collagenase type I  in α-MEM for 30 minutes at 37℃ with gentle shaking. Cells were filtered through a 70 μm cell strainer and centrifuged to obtain the cell pellet, then re-suspended with FACS buffer to receive the cell suspension. The Gli1+ progenitors were sorted using a BD Arial III according to the presence or absence of PE signal using a C57BL/6J mouse as a negtive control. Sequencing libraries constructed with reagents from a Chromium Single Cell 3' v2 reagent kit (10X Genomics) according to the manufacturer's instructions.