Evaluation of the in vitro effects of irisin and leptin on human ovarian granulosa cells – PCR array data (female infertility panel)

Published: 12 November 2021| Version 1 | DOI: 10.17632/gr3dg36nzx.1
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Description

Human ovarian granulosa cells were purified from follicular fluid samples obtained from patients undergoing in vitro fertilization procedure by using Percoll PLUS reagent (GE Healthcare, Cat. # 17-5445-02). Cells were grown in DMEM/F12 (50:50) medium (Corning, Cat. # 10-092-CM) supplemented with 10% FBS (VWR, Cat. # 89510-186) and antibiotic/antimycotic mixture (MP Biomedicals, Cat. # 1674049). For experiment, 0.3 x 10(6) cells were seeded in 6-well plates and the following day the cell culture medium was replaced with serum-free medium supplemented with 500 ng/ml of irisin (Enzo, Cat. # ADI-908-307-0010) or 50 ng/ml of leptin (BioVision, Cat. # 4366-02) for 24 hours. After completing the experiment, cells were washed two times with PBS (Corning, Cat. # 46-013-CM) and total RNA was extracted using TRIzol reagent (Ambion, 15596018). RNA was quantified using NanoDrop One spectrophotometer (Thermo Fisher Scientific) and samples were normalized to 1 mg/ml concentration, then converted to cDNA using qScript cDNA SuperMix (Quantabio, Cat. # 95048). PCR array was performed using RT2 Profiler PCR Array – Human Female Infertility (Qiagen, Cat. # 330231, GeneGlobe ID PAHS-164Z) and QuantStudio 3 Real-Time PCR System (Thermo Fisher Scientific) following manufacturer’s protocol. PCR array data were calculated using GeneGlobe Data Analysis Center (https://geneglobe.qiagen.com/us/analyze).

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Institutions

Lenox Hill Hospital, Donald and Barbara Zucker School of Medicine at Hofstra/Northwell, Northwell Health

Categories

Cytokines, Female Reproduction, Leptin, Infertility, Data Array, Signal Transduction, Adipokine, Leptin Treatment, Quantitative Reverse Transcription Polymerase Chain Reaction

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