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Updated: Nov 6, 2025
The Production of Pluripotent Stem Cells from Mouse Amniotic Fluid Cells Using a Transposon System
Published on: February 28, 2017
miR-351-3p promotes rat amniotic fluid-derived mesenchymal stromal cell proliferation via targeting the coding
Jieting Huang1,2,3, Qi Zhao4, Xiaowei Wei3
1BaYi Children's Hospital, Seventh Medical Center of Chinese PLA General Hospital, Beijing, People's Republic of China.
Abstract:
Amniotic fluid-derived mesenchymal stromal cells (AFMSCs) present different features, depending on the isolation timing and culture conditions. The lack of uniform experimental standards hinders the comparison of results from different studies on AFMSCs. Moreover, understanding the molecular mechanisms that underlie the features of AFMSCs isolated at different embryonic developmental stages might allow the obtention of more viable and highly proliferative AFMSCs through genetic modification. We isolated AFMSCs from pregnant rats at embryonic day (E)12, E15, E18, and E21 and compared their cell proliferation capacity and transcriptome. The cell counting kit-8 assay and RNA sequencing revealed that E12 and E15 AFMSCs showed different characteristics from E18 and E21 AFMSCs. Therefore, AFMSCs were divided into two groups: early (E12 and E15) and late (E18 and E21) pregnancy-stage groups. Next, we screened the gene/microRNA pair Abca4/miR-351-3p that was related to cell proliferation. Abca4 knockdown/overexpression suggested that this gene represses the proliferation of AFMSCs, which is a newly discovered function of this gene. Finally, dual luciferase reporter gene assays confirmed that miR-351-3p targeted the coding sequence of Abca4 and regulated AFMSC proliferation. miR-351-3p promotes AFMSC proliferation via targeting the coding sequence of Abca4. Our findings provide a molecular foundation for further research for obtaining AFMSCs with a higher proliferation capacity.
Insights
Amniotic fluid-derived mesenchymal stromal cells (AFMSCs) from early pregnancy stages exhibit distinct characteristics. A newly identified mechanism shows miR-351-3p promotes AFMSC proliferation by targeting Abca4.
Area of Science:
- Stem Cell Biology
- Developmental Biology
- Molecular Genetics
Background:
- Amniotic fluid-derived mesenchymal stromal cells (AFMSCs) exhibit variable features influenced by isolation and culture methods.
- Standardization is lacking for AFMSC research, hindering result comparisons.
- Understanding molecular drivers of AFMSC characteristics at different developmental stages can guide strategies for enhanced proliferation and viability.
Purpose of the Study:
- To compare the proliferation capacity and transcriptomes of AFMSCs isolated at different embryonic developmental stages in rats.
- To identify molecular mechanisms, specifically gene/microRNA interactions, regulating AFMSC proliferation.
- To investigate the role of the Abca4/miR-351-3p axis in controlling AFMSC proliferation.
Main Methods:
- Isolation and culture of AFMSCs from rat embryos at embryonic days (E)12, E15, E18, and E21.
- Cell counting kit-8 assay for proliferation assessment and RNA sequencing for transcriptome analysis.
- Gene knockdown/overexpression studies for Abca4 and dual luciferase reporter gene assays for miR-351-3p-Abca4 interaction validation.
Main Results:
- AFMSCs were categorized into early (E12, E15) and late (E18, E21) pregnancy-stage groups based on distinct characteristics.
- The gene/microRNA pair Abca4/miR-351-3p was identified as a regulator of AFMSC proliferation.
- Abca4 was found to repress AFMSC proliferation, while miR-351-3p promotes it by targeting Abca4's coding sequence.
Conclusions:
- AFMSCs isolated at different embryonic stages possess distinct properties.
- The miR-351-3p targeting of Abca4 represents a novel mechanism regulating AFMSC proliferation.
- These findings offer a molecular basis for developing strategies to enhance AFMSC proliferation capacity.
