Comparative bulk RNAseq analysis of CXCL12-abundant reticular (CAR) cells, fetal perichondrium-derived CAR cells and chondrocyte-derived CAR cells.

We revealed the unique molecular signature characterizing fetal perichondrium-derived and chondrocyte-derived CAR cells GFP+tdTomato+ cells were FACS-isolated from postnatal day 21 (P21) femurs of Cxcl12-GFP; Hes1-creER; R26R-tdTomato and Cxcl12-GFP; Fgfr3-iCreER; R26R-tdTomato mice, both pulsed with a single dose of tamoxifen at embryonic day 12.5 (E12.5). mRNA were amplifed by SMARTer mRNA-seq kit (Clonetech/Takara) before deep-sequenced by Illumina HiSeq4000.

本研究揭示了胎儿软骨膜来源及软骨细胞来源的CAR细胞的独特分子特征。GFP与tdTomato双阳性细胞通过荧光激活细胞分选术（Fluorescence-Activated Cell Sorting, FACS），从Cxcl12-GFP; Hes1-creER; R26R-tdTomato与Cxcl12-GFP; Fgfr3-iCreER; R26R-tdTomato小鼠的出生后第21天（P21）股骨中分离得到；上述两种小鼠均于胚胎发育第12.5天（E12.5）接受单次他莫昔芬诱导处理。mRNA经SMARTer mRNA测序试剂盒（Clonetech/Takara）扩增后，采用Illumina HiSeq4000平台完成深度测序。