Illumina HiSeq 2000 sequencing; GSM2581177: 11E_1_epithelium_LH2; Homo sapiens; RNA-Seq

11E_1_epithelium_LH2

Cell-type specific RNA-seq is a powerful approach for unravelling molecular processes of endometrial receptivity, and to detect novel sensitive biomarkers of receptivity. Overall design: 16 paired endometrial tissue samples from pre-receptive (defined as LH2) and receptive phase endometria (defined as LH8) from Estonia (defined as E) and Spain (defined as S) were collected. CD9-positive epithelial cells (defined as epithelium) and CD13-positive stromal cells (defined as stroma) were isolated with fluorescent activated cell sorting (FACS) and full transcriptome analysis was performed by RNA-seq.

Endometrial cell-type specific RNA-seq

Submitted on 08-NOV-2017

Endometrial biopsies were collected with endometrial suction Pipelle catheter (Laboratoire CCD, France). Briefly the biopsied tissue samples were dissociated, endometrial stromal cells were stained with fluorocrome-conjugated mouse anti-human CD13 monoclonal antibody (clone TUK1, R-Phycoerythrin, Invitrogen, USA), epithelial cells were stained simultaneously with fluorocrome-conjugated mouse anti-human CD9 monoclonal antibody (clone MEM-61, FITC, Novus Biologicals, USA) and all dead cells were stained with DAPI (Invitrogen, USA).CD9 or CD13 positive and DAPI negative (alive) cells were sorted into a TRIzol Reagent and RNA were extracted immediately after the lysis of cells. Bulk-RNA full transcriptome analysis of FACS sorted endometrial cells was performed by the RNA-seq method, following the single-cell tagged reverse transcription (STRT) protocol with modifications (KrjutÅ¡kov et al., 2016). 10 ng of high-quality input of cell-type specific RNA was converted to cDNA and amplified to form an Illumina-compatible library. STRT (detailed protocol in PubMed ID 26874359), RNA-seq

Endometrial cell-type specific RNA-seq

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