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Updated: Feb 12, 2026

Isolation and Differentiation of Primary White and Brown Preadipocytes from Newborn Mice
Published on: January 25, 2021
Identification of differentially expressed microRNAs during preadipocyte differentiation in Chinese crested duck
Shasha Wang1, Yang Zhang1, Xiaoya Yuan1
1Key Laboratory of Animal Genetics and Breeding and Molecular Design of Jiangsu Province, College of Animal Science and Technology, Yangzhou University, Yangzhou, Jiangsu, China.
Abstract:
MicroRNAs (miRNAs) are considered key players in the regulation of a broad range of biological processes. Specifically, miRNAs have been reported to play an important role in the process of adipogenesis. In this study, we constructed a model of adipogenesis by isolating preadipocytes (WCC) derived from adipose tissue and preadipocytes after 72 h differentiation (WCT) in vitro. Deep sequencing of miRNAs expressed in WCT and WCC cells was conducted; we identified 105 differentially expressed miRNA candidates (fifty up-regulated and fifty-five down-regulated). Among them, twelve were novel miRNAs, and ninety-three were previously known miRNAs. Furthermore, seven miRNAs were selected for expression confirmation by reverse transcription quantitative PCR (RT-qPCR); the results showed that the differential expression of miRNAs between the two groups was consistent with our sequencing results. Of them, miR-223, miR-184-3p, and miR-10b-5 showed a strong correlation to adipogenesis. Using target prediction, we predicted that the 105 differentially expressed miRNAs targeted 4155 unique mRNAs. The prediction of targets of differentially expressed miRNAs revealed that the miRNAs participated in the regulation of multiple adipogenesis-related signalling pathways, including the peroxisome proliferator-activated receptor (PPAR) signalling pathway, insulin signalling pathway, fatty acid biosynthesis, and fatty acid degradation. Overall, our findings provide a background for further research into miRNAs and lay a foundation for the prediction and analysis of miRNAs related to adipogenesis.
Insights
MicroRNAs regulate adipogenesis, with 105 differentially expressed candidates identified. Key miRNAs like miR-223 correlate with fat cell development, impacting crucial metabolic pathways.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- MicroRNAs (miRNAs) are crucial regulators of biological processes, including adipogenesis.
- Understanding miRNA roles in adipogenesis is vital for metabolic research.
Purpose of the Study:
- To identify differentially expressed miRNAs during adipogenesis.
- To investigate the regulatory roles of these miRNAs in adipogenesis-related pathways.
Main Methods:
- Isolation and in vitro differentiation of preadipocytes (WCC and WCT).
- Deep sequencing of miRNA expression profiles.
- Reverse transcription quantitative PCR (RT-qPCR) for validation.
- Bioinformatic target prediction analysis.
Main Results:
- Identified 105 differentially expressed miRNA candidates (50 upregulated, 55 downregulated).
- Confirmed differential expression of selected miRNAs, including miR-223, miR-184-3p, and miR-10b-5, which strongly correlate with adipogenesis.
- Predicted 4155 unique mRNA targets for the identified miRNAs.
- Revealed involvement of these miRNAs in PPAR, insulin signaling, fatty acid biosynthesis, and degradation pathways.
Conclusions:
- Established a comprehensive profile of differentially expressed miRNAs during adipogenesis.
- Highlighted specific miRNAs (e.g., miR-223) as key regulators of adipogenesis.
- Provided a foundation for further research into miRNA-mediated control of adipogenesis and related metabolic functions.
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