miR-351-3p Promotes Rat Amniotic Fluid-Derived Mesenchymal Stromal Cell Proliferation via Targeting the Coding Sequence of Abca4

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.

Applications of human amniotic fluid stem cells in wound healing

“…Differentiation of AFSCs is regulated by DNA methylation, histone modification, and the expression of small non-coding RNAs, [28] most notably the miR-351-3p, a gene that targets Abca4 (ATP-binding cassette transporter A4, Abca4) and regulates the proliferation and migration of AFSCs by targeting the 3 0 -UTR (untranslated region, UTR) of Chrdl1 (chordin-like 1, Chrdl1) and downregulating its expression. [29,30] Reports have shown that AFSCs grow well in a serum-free medium, and the proliferation efficiency was higher in the AmnioMAX medium than in Dulbecco's modified Eagle medium. [12,31] The cells have a doubling time of about 36 h, can proliferate 250 times in vitro without chromosome loss, [3] and maintain stable telomere lengths over several generations.…”

“…The phenotypic characteristics of AFSCs vary with the stage of pregnancy, [3] and epigenetic modifications guide AFSC differentiation into multiple lineages, playing a vital role in stem cell fate determination. Differentiation of AFSCs is regulated by DNA methylation, histone modification, and the expression of small non-coding RNAs, [28] most notably the miR-351-3p, a gene that targets Abca4 (ATP-binding cassette transporter A4, Abca4) and regulates the proliferation and migration of AFSCs by targeting the 3′-UTR (untranslated region, UTR) of Chrdl1 (chordin-like 1,Chrdl1) and downregulating its expression [29,30] …”

Roles of mesenchymal stem cells and exosomes in interstitial cystitis/bladder pain syndrome

“…MSCs can be isolated from different adult tissues such as umbilical cord blood, bone marrow, adipose tissue, placenta, amniotic fluid, urine and other tissues. 7 , 8 , 9 , 10 , 11 They express low levels of major histocompatibility complex (MHC) class I molecules and MHC class II molecules, which enable them to have low immunogenicity. 12 In vitro, MSCs can differentiate into a range of cell types, including adipocytes, osteocytes, hepatocytes, chondrocytes, muscle, vascular smooth muscle cells and other connective tissues.…”