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Updated: Jun 29, 2025
Gene Expression Analyses in Human Follicles
Published on: February 17, 2023
Evaluation of the expression level of microRNA-21, microRNA-15a, microRNA-372 in human follicular fluid stem
Ghasem Saki1,2, Kousar Shahrooie1,2, Mahin Taheri Moghadam1,2
1Cellular and Molecular Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.
Objective:
Today, researchers have succeeded in achieving oocyte-like cells through the in vitro differentiation of stem cells. MicroRNAs are key regulators of oocyte development. In this study we decided to evaluate the expression pattern of microRNA-21, microRNA-15a, and microRNA-372 in oocyte-like cells, to determine the maturation stage of oocyte-like cells.
Methods:
Human follicular fluid samples were collected and centrifuged, and their cells were divided into 3 groups; day 7 as control group, days 14 and 21. During this period, the cells were evaluated for their morphological appearance and viability by inverted microscopy. RNA isolation was performed and cDNA was reversely transcribed by specific stem-loop RT primers. Real-time RT-PCR was used to detect microRNA expression.
Results:
The relative expression of microRNA-21 and microRNA-15a on day 21 was significantly down-regulated compared to the control group (day 7), but microRNA-372 did not show a significant difference. Also, on day 14 compared to the control group (day 7), microRNA-21 did not show a significant difference; but microRNA-15a and microRNA-372 were significantly down-regulated. MicroRNA-21 and microRNA-15a on day 21 compared to day 14 revealed down-regulated levels, but microRNA-372 revealed up-regulated levels.
Conclusions:
Our results showed significant decreases in the expression of microRNA-21 and microRNA-15a in oocyte-like cells, as well as in oocytes, which may lead to cytoplasmic maturation, germinal vesicle break down and the completion of meiosis І. In addition, down-regulation expression of microRNA-372 maybe a confirmation that mesenchymal stem cells have differentiated into germ cells, and these cells were differentiated into oocyte-like cells.
Insights
Researchers studied microRNA-21, microRNA-15a, and microRNA-372 expression in oocyte-like cells. Significant down-regulation of microRNA-21 and microRNA-15a suggests successful stem cell differentiation and oocyte maturation.
Area of Science:
- Stem cell biology
- Reproductive biology
- Molecular genetics
Background:
- Stem cell differentiation holds promise for generating oocyte-like cells in vitro.
- MicroRNAs (miRNAs) are critical regulators of oocyte development and maturation.
- Understanding miRNA expression patterns is key to assessing the developmental stage of in vitro-derived oocyte-like cells.
Purpose of the Study:
- To investigate the expression patterns of microRNA-21, microRNA-15a, and microRNA-372 in oocyte-like cells derived from stem cells.
- To correlate miRNA expression levels with the maturation stage of these oocyte-like cells.
Main Methods:
- Human follicular fluid cells were cultured and divided into control (day 7) and experimental groups (days 14 and 21).
- Morphological assessment and viability testing were performed using inverted microscopy.
- RNA isolation, stem-loop RT-primer-based cDNA synthesis, and real-time RT-PCR were employed to quantify miRNA expression.
Main Results:
- MicroRNA-21 and microRNA-15a showed significant down-regulation on day 21 compared to day 7.
- MicroRNA-15a and microRNA-372 were significantly down-regulated on day 14 versus day 7.
- MicroRNA-372 levels increased from day 14 to day 21, while microRNA-21 and microRNA-15a decreased.
Conclusions:
- Significant decreases in microRNA-21 and microRNA-15a expression in oocyte-like cells correlate with potential cytoplasmic maturation and completion of meiosis I.
- The observed down-regulation of microRNA-372 may indicate successful differentiation of mesenchymal stem cells into germ cells and subsequently into oocyte-like cells.
