Hossein Faghih, Maryam Khani, Mehdi Shamsara, Hossein Taghizadeh, Arash Javeri, Masoumeh Fakhr Taha
{"title":"通过miR-302/367簇过表达增强人脂肪组织源性干细胞的多能性和成脂分化","authors":"Hossein Faghih, Maryam Khani, Mehdi Shamsara, Hossein Taghizadeh, Arash Javeri, Masoumeh Fakhr Taha","doi":"10.1007/s12195-025-00856-z","DOIUrl":null,"url":null,"abstract":"<p><strong>Purpose: </strong>Human adipose tissue-derived stem cells (hADSCs) have emerged as a promising source of cells for tissue engineering and regenerative medicine. However, their differentiation potential is restricted and requires enhancements. This study explores the reprogramming of hADSCs through exogenous induction of the miR-302/367 cluster.</p><p><strong>Methods: </strong>Human ADSCs were transfected with the mock or miR-302/367 cluster-expressing vectors. One week after transfection, expression levels of several pluripotency-related genes, epithelial-to-mesenchymal (EMT) markers, and mechanistic target of rapamycin kinase (mTOR) signaling factors were assessed by qPCR and western blot. Additionally, the influence of miR-302/367 cluster overexpression on the proliferation and adipogenic differentiation of the ADSCs was evaluated.</p><p><strong>Results: </strong>One week after transfection, the expression of several pluripotency-related genes and epithelial markers was significantly upregulated, while mesenchymal markers were downregulated in the miR-302/367-transfected cells compared with the mock group. Additionally, the levels of several mTOR signaling factors were reduced in the miR-302/367-transfected ADSCs. Flow cytometry analysis showed a decrease in the abundance of ADSCs in the S phase and an increase in the population of cells in the G1 phase of the cell cycle. Moreover, the adipogenic differentiation of the miR-302/367-transfected cells was significantly enhanced.</p><p><strong>Conclusion: </strong>The overexpression of the miR-302/367 cluster directed the ADSCs towards a more pluripotent state and promoted their adipogenic potential. However, miR-302/367 overexpression diminished the proliferative capacity of hADSCs, which warrants a comprehensive investigation. Further evaluations are needed to fully elucidate the differentiation potential and regenerative capacity of the ADSCs reprogrammed by the miR-302/367 cluster before any clinical application.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at 10.1007/s12195-025-00856-z.</p>","PeriodicalId":9687,"journal":{"name":"Cellular and molecular bioengineering","volume":"18 3-4","pages":"297-309"},"PeriodicalIF":5.0000,"publicationDate":"2025-08-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12436253/pdf/","citationCount":"0","resultStr":"{\"title\":\"Enhancing Pluripotency and Adipogenic Differentiation in Human Adipose Tissue-Derived Stem Cells Through miR-302/367 Cluster Overexpression.\",\"authors\":\"Hossein Faghih, Maryam Khani, Mehdi Shamsara, Hossein Taghizadeh, Arash Javeri, Masoumeh Fakhr Taha\",\"doi\":\"10.1007/s12195-025-00856-z\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Purpose: </strong>Human adipose tissue-derived stem cells (hADSCs) have emerged as a promising source of cells for tissue engineering and regenerative medicine. However, their differentiation potential is restricted and requires enhancements. This study explores the reprogramming of hADSCs through exogenous induction of the miR-302/367 cluster.</p><p><strong>Methods: </strong>Human ADSCs were transfected with the mock or miR-302/367 cluster-expressing vectors. One week after transfection, expression levels of several pluripotency-related genes, epithelial-to-mesenchymal (EMT) markers, and mechanistic target of rapamycin kinase (mTOR) signaling factors were assessed by qPCR and western blot. Additionally, the influence of miR-302/367 cluster overexpression on the proliferation and adipogenic differentiation of the ADSCs was evaluated.</p><p><strong>Results: </strong>One week after transfection, the expression of several pluripotency-related genes and epithelial markers was significantly upregulated, while mesenchymal markers were downregulated in the miR-302/367-transfected cells compared with the mock group. Additionally, the levels of several mTOR signaling factors were reduced in the miR-302/367-transfected ADSCs. Flow cytometry analysis showed a decrease in the abundance of ADSCs in the S phase and an increase in the population of cells in the G1 phase of the cell cycle. Moreover, the adipogenic differentiation of the miR-302/367-transfected cells was significantly enhanced.</p><p><strong>Conclusion: </strong>The overexpression of the miR-302/367 cluster directed the ADSCs towards a more pluripotent state and promoted their adipogenic potential. However, miR-302/367 overexpression diminished the proliferative capacity of hADSCs, which warrants a comprehensive investigation. Further evaluations are needed to fully elucidate the differentiation potential and regenerative capacity of the ADSCs reprogrammed by the miR-302/367 cluster before any clinical application.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at 10.1007/s12195-025-00856-z.</p>\",\"PeriodicalId\":9687,\"journal\":{\"name\":\"Cellular and molecular bioengineering\",\"volume\":\"18 3-4\",\"pages\":\"297-309\"},\"PeriodicalIF\":5.0000,\"publicationDate\":\"2025-08-11\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12436253/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Cellular and molecular bioengineering\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://doi.org/10.1007/s12195-025-00856-z\",\"RegionNum\":4,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2025/8/1 0:00:00\",\"PubModel\":\"eCollection\",\"JCR\":\"Q3\",\"JCRName\":\"BIOPHYSICS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Cellular and molecular bioengineering","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1007/s12195-025-00856-z","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/8/1 0:00:00","PubModel":"eCollection","JCR":"Q3","JCRName":"BIOPHYSICS","Score":null,"Total":0}
Enhancing Pluripotency and Adipogenic Differentiation in Human Adipose Tissue-Derived Stem Cells Through miR-302/367 Cluster Overexpression.
Purpose: Human adipose tissue-derived stem cells (hADSCs) have emerged as a promising source of cells for tissue engineering and regenerative medicine. However, their differentiation potential is restricted and requires enhancements. This study explores the reprogramming of hADSCs through exogenous induction of the miR-302/367 cluster.
Methods: Human ADSCs were transfected with the mock or miR-302/367 cluster-expressing vectors. One week after transfection, expression levels of several pluripotency-related genes, epithelial-to-mesenchymal (EMT) markers, and mechanistic target of rapamycin kinase (mTOR) signaling factors were assessed by qPCR and western blot. Additionally, the influence of miR-302/367 cluster overexpression on the proliferation and adipogenic differentiation of the ADSCs was evaluated.
Results: One week after transfection, the expression of several pluripotency-related genes and epithelial markers was significantly upregulated, while mesenchymal markers were downregulated in the miR-302/367-transfected cells compared with the mock group. Additionally, the levels of several mTOR signaling factors were reduced in the miR-302/367-transfected ADSCs. Flow cytometry analysis showed a decrease in the abundance of ADSCs in the S phase and an increase in the population of cells in the G1 phase of the cell cycle. Moreover, the adipogenic differentiation of the miR-302/367-transfected cells was significantly enhanced.
Conclusion: The overexpression of the miR-302/367 cluster directed the ADSCs towards a more pluripotent state and promoted their adipogenic potential. However, miR-302/367 overexpression diminished the proliferative capacity of hADSCs, which warrants a comprehensive investigation. Further evaluations are needed to fully elucidate the differentiation potential and regenerative capacity of the ADSCs reprogrammed by the miR-302/367 cluster before any clinical application.
Supplementary information: The online version contains supplementary material available at 10.1007/s12195-025-00856-z.
期刊介绍：
The field of cellular and molecular bioengineering seeks to understand, so that we may ultimately control, the mechanical, chemical, and electrical processes of the cell. A key challenge in improving human health is to understand how cellular behavior arises from molecular-level interactions. CMBE, an official journal of the Biomedical Engineering Society, publishes original research and review papers in the following seven general areas:
Molecular: DNA-protein/RNA-protein interactions, protein folding and function, protein-protein and receptor-ligand interactions, lipids, polysaccharides, molecular motors, and the biophysics of macromolecules that function as therapeutics or engineered matrices, for example.
Cellular: Studies of how cells sense physicochemical events surrounding and within cells, and how cells transduce these events into biological responses. Specific cell processes of interest include cell growth, differentiation, migration, signal transduction, protein secretion and transport, gene expression and regulation, and cell-matrix interactions.
Mechanobiology: The mechanical properties of cells and biomolecules, cellular/molecular force generation and adhesion, the response of cells to their mechanical microenvironment, and mechanotransduction in response to various physical forces such as fluid shear stress.
Nanomedicine: The engineering of nanoparticles for advanced drug delivery and molecular imaging applications, with particular focus on the interaction of such particles with living cells. Also, the application of nanostructured materials to control the behavior of cells and biomolecules.
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