ArticleBeijing da xue xue bao. Yi xue ban = Journal of Peking University. Health sciences2026
[A new method for extracting adult mouse cardiac fibroblasts more efficiently and stably].
Article in Beijing da xue xue bao. Yi xue ban = Journal of Peking University. Health sciences, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
objectiveCardiac fibroblasts (CFs) play a central role in myocardial remodeling and fibrosis. Efficient isolation of CFs is a prerequisite for investigating related mechanisms. However, current methods for isolating primary adult mouse CFs suffer from prolonged processing time, low yield, and poor viability. This study aims to establish a rapid, high-yield, and stable isolation protocol for adult mouse CFs by optimizing the synergistic effect of enzymatic digestion and mechanical dissociation parameters.
methodsUsing the gentleMACS
resultsOmitting any component of the digestion solution (collagenase Ⅱ/Ⅳ, trypsin or DNaseⅠ), significantly prolonged extraction time and reduced cell yield. In contrast, the optimized protocol outperformed the commercial kit, reducing digestion time by 32.2 min and significantly increasing cell yield, and with comparable obtained CFs purity. After TGF-β1 stimulation, CFs exhibited enhanced proliferative capacity and upregulated expression of α-smooth muscle actin (α-SMA), collagen type Ⅰ (ColⅠ), and fibronectin (FN), confirming the differentiation potential of CFs isolated via the optimized method.
conclusionThis study systematically optimized an enzymatic digestion method combining collagenase, trypsin, and nuclease in conjunction with mechanical dissociation using a tissue dissociator, leading to the efficient and stable isolation of adult mouse CFs. By fine-tuning enzyme concentrations and digestion conditions, we successfully reduced processing time, improved cell yield, and enhanced cell viability compared with conventional isolation methods. These findings validate the physiological relevance of the isolated CFs and demonstrate that the optimized protocol provides a reliable and reproducible method for studying myocardial fibrosis and remodeling. This protocol can serve as a valuable tool for researchers investigating CFs biology and its role in cardiovascular diseases.
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42287063PMC13268823What OpenQuestion holds
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