GSM5899025: Testicular Microenvironment_B04_Lep 10; Homo sapiens; RNA-Seq
The protocol for LSC isolation has been described previously (12). Briefly, interstitial cells from the testes were dissociated from the seminiferous tubules by treatment with 1 mg/ml trypsin followed by collagenase (collagenase D; Roche Molecular Biochemicals) treatment in DMEM for 10 min at 34°C with shaking.  The separated cells were filtered through two layers of 70-μm pore-size nylon mesh, centrifuged at 250 × g, and re-suspended in DMEM. The cells were washed with PBS, centrifuged at 250 × g, re-suspended in phenol red-free 1:1 DMEM: F12 and plated for cell culture. Truseq standard Total RNA Library Prep Libraries were prepared according to Illumina's instructions accompanying the Sample Kit (Truseq standard Total RNA Library Prep).