Light/Dark Environmental Cycle Imposes a Daily Profile in the Expression of microRNAs in Rat CD133(+) Cells
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Updated: Oct 24, 2026
Differentiation of a Human Neural Stem Cell Line on Three Dimensional Cultures, Analysis of MicroRNA and Putative Target Genes
Published on: April 12, 2015
Marina Marçola1, Camila M Lopes-Ramos2, Eliana P Pereira1
1Department of Physiology, Laboratory of Chronopharmacology, Institute of Bioscience, University of São Paulo, São Paulo City, São Paulo, Brazil.
Abstract:
The phenotype of primary cells in culture varies according to the donor environmental condition. We recently showed that the time of the day imposes a molecular program linked to the inflammatory response that is heritable in culture. Here we investigated whether microRNAs (miRNAs) would show differential expression according to the time when cells were obtained, namely daytime or nighttime. Cells obtained from explants of cremaster muscle and cultivated until confluence (∼20 days) presented high CD133 expression. Global miRNA expression analysis was performed through deep sequencing in order to compare both cultured cells. A total of 504 mature miRNAs were identified, with a specific miRNA signature being associated to the light versus dark phase of a circadian cycle. miR-1249 and miR-129-2-3p were highly expressed in daytime cells, while miR-182, miR-96-5p, miR-146a-3p, miR-146a-5p, and miR-223-3p were highly expressed in nighttime cells. Nighttime cells are regulated for programs involved in cell processes and development, as well as in the inflammation, cell differentiation and maturation; while daytime cells express miRNAs that control stemness and cytoskeleton remodeling. In summary, the time of the day imposes a differential profile regarding to miRNA signature on CD133(+) cells in culture. Understanding this daily profile in the phenotype of cultured cells is highly relevant for clinical outputs, including cellular therapy approaches. J. Cell. Physiol. 231: 1953-1963, 2016. © 2016 Wiley Periodicals, Inc.
Insights
The time of day influences microRNA (miRNA) expression in cultured CD133(+) cells. This daily miRNA signature impacts cell function and is relevant for cellular therapies.
Area of Science:
- Cell Biology
- Molecular Biology
- Circadian Biology
Background:
- Primary cell phenotype in culture is influenced by donor conditions.
- The time of day can impose heritable molecular programs related to inflammation.
- MicroRNAs (miRNAs) are key regulators of gene expression.
Purpose of the Study:
- To investigate differential microRNA expression in cultured cells based on collection time (daytime vs. nighttime).
- To identify specific miRNA signatures associated with circadian phases in cultured cells.
- To understand the functional implications of time-dependent miRNA profiles in CD133(+) cells.
Main Methods:
- Primary cremaster muscle cells were cultured until confluence.
- High CD133 expression was confirmed in cultured cells.
- Global miRNA expression analysis was performed using deep sequencing.
Main Results:
- A total of 504 mature miRNAs were identified.
- Distinct miRNA signatures were observed between daytime and nighttime cells, linked to circadian cycles.
- Specific miRNAs (e.g., miR-1249, miR-129-2-3p) were enriched in daytime cells, while others (e.g., miR-182, miR-96-5p) were enriched in nighttime cells.
- Daytime cells showed miRNAs regulating stemness and cytoskeleton remodeling.
- Nighttime cells exhibited miRNAs involved in cell processes, development, inflammation, and differentiation.
Conclusions:
- The time of day significantly impacts the miRNA signature of cultured CD133(+) cells.
- Understanding these daily miRNA profiles is crucial for optimizing cell culture and clinical applications, particularly in cellular therapy.
- Circadian rhythm influences cellular phenotype at the miRNA level in vitro.
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