AI Article Synopsis
- The study presents a method to isolate specific cell types, like pluripotent stem cell (PSC)-derived populations, using endogenous miRNA activities instead of relying solely on known cell surface antigens.
- By utilizing synthetic mRNAs encoding fluorescent proteins that respond to miRNAs from target cells, the method can efficiently purify cardiomyocytes and other cell types.
- This innovative miRNA switch technique proves effective for various cell types, including endothelial cells and insulin-producing cells, providing an alternative isolation strategy when traditional methods are not available.
Article Abstract
Isolation of specific cell types, including pluripotent stem cell (PSC)-derived populations, is frequently accomplished using cell surface antigens expressed by the cells of interest. However, specific antigens for many cell types have not been identified, making their isolation difficult. Here, we describe an efficient method for purifying cells based on endogenous miRNA activity. We designed synthetic mRNAs encoding a fluorescent protein tagged with sequences targeted by miRNAs expressed by the cells of interest. These miRNA switches control their translation levels by sensing miRNA activities. Several miRNA switches (miR-1-, miR-208a-, and miR-499a-5p-switches) efficiently purified cardiomyocytes differentiated from human PSCs, and switches encoding the apoptosis inducer Bim enriched for cardiomyocytes without cell sorting. This approach is generally applicable, as miR-126-, miR-122-5p-, and miR-375-switches purified endothelial cells, hepatocytes, and insulin-producing cells differentiated from hPSCs, respectively. Thus, miRNA switches can purify cell populations for which other isolation strategies are unavailable.
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| http://dx.doi.org/10.1016/j.stem.2015.04.005 | DOI Listing |
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