Comparison of Cumulus Cells Preparation among Diminished Ovarian Reserved and Normal Ovarian Reserved Women for RNA Extraction and Review of Literature

Our study aims to consolidate appropriate cumulus cells (CCs) collection methods by comparing sample handling obtained from diminished ovarian reserved (DOR) and normal ovarian reserved (NOR) women, aiming for a good yield of ribonucleic acid (RNA) for better gene expression analysis. We compared the sample collection by mechanical versus enzymatic method, standard rotor-stator homogeniser (RSH) versus disposable easy grind homogeniser for sample preparation technique and subsequently the cDNA synthesis and qPCR for final evaluation. We also conducted a systematic review to consolidate our findings with current evidence of RNA extraction technique. The optimal strategy was via mechanical denudation without an enzymatic process, followed by RNA stabiliser prior to RNA extraction. Subsequently, utilising a manual pressure control with filter, is paramount for a pure and homogenised sample and comparable for both NOR and DOR women CCs. These sample preparation and extraction techniques yielded an optimum RNA concentration and successful gene expression analysis. Our review also added value as a current strategy for optimising RNA extraction in CCs for experimental studies. Our findings contribute to formulating a better strategy for optimising RNA extraction in human CCs for transcriptomic studies.