GSM7690206
Cells were isolated from the tibia and femur and crushed once, using mortar and pestle, then cut into small pieces with scissors. The tissue was subjected to three rounds of enzymatic digestion with 0.2%collagenaseat 37°C under agitation for 30min each. Cells were filtered through a 70-μm strainer and centrifuged at 1500 rpm for 5 minutes at 4°C.  Red blood cells were lysed using NH4Cl (StemCell Technologies, Vancouver, Canada) for 10 min on ice, washed with staining media (HBSS (Thermo Fisher Scientific, Waltham, MA, USA) containing 2% fetal bovine serum (Life Technologies: 10437-028), 1% HEPES (10 mM) (Thermo Fisher Scientific) and 1% penicillin-streptomycin (Thermo Fisher Scientific) and resuspended in HBSS for subsequent analyses RNA was isolated from cells using RNeasy Kit (Qiagen, Germantown, MD, USA) according to manufacturer's instruction RNA-seq, Low input RNA (10 pg) library prep (SMARTer or Nugen method)