{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,19]],"date-time":"2026-02-19T04:30:59Z","timestamp":1771475459990,"version":"3.50.1"},"reference-count":52,"publisher":"MDPI AG","issue":"23","license":[{"start":{"date-parts":[[2024,11,23]],"date-time":"2024-11-23T00:00:00Z","timestamp":1732320000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"WISS 2025 project \u2018IDA-Lab Salzburg\u2019","award":["20204-WISS\/225\/197-2019"],"award-info":[{"award-number":["20204-WISS\/225\/197-2019"]}]},{"name":"WISS 2025 project \u2018IDA-Lab Salzburg\u2019","award":["20102-F1901166-KZP"],"award-info":[{"award-number":["20102-F1901166-KZP"]}]},{"name":"WISS 2025 project \u2018IDA-Lab Salzburg\u2019","award":["20204-WISS\/263\/6-6022"],"award-info":[{"award-number":["20204-WISS\/263\/6-6022"]}]},{"name":"Land Salzburg","award":["20204-WISS\/225\/197-2019"],"award-info":[{"award-number":["20204-WISS\/225\/197-2019"]}]},{"name":"Land Salzburg","award":["20102-F1901166-KZP"],"award-info":[{"award-number":["20102-F1901166-KZP"]}]},{"name":"Land Salzburg","award":["20204-WISS\/263\/6-6022"],"award-info":[{"award-number":["20204-WISS\/263\/6-6022"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>In regenerative medicine, stromal cells are supposed to play an important role by modulating immune responses and differentiating into various tissue types. The aim of this study was to investigate the influence of heparin, frequently used as an anticoagulant in human platelet lysate (HPL)-supplemented cell cultures, on the expression of non-coding RNA species, particularly microRNAs (miRNA), which are pivotal regulators of gene expression. Through genomic analysis and quantitative RT-PCR, we assessed the differential impact of heparin on miRNA expression in various stromal cell types, derived from human bone marrow, umbilical cord and white adipose tissue. Our results demonstrate that heparin significantly alters miRNA expression, with distinct up- and downregulation patterns depending on the original tissue source of human stromal cells. Furthermore, our analyses indicate that these heparin-induced alterations in miRNA expression profiles influence critical cellular processes, including proliferation, apoptosis and differentiation. In conclusion, our study highlights that heparin not only fulfills its primary role as an efficient anticoagulant but can also modulate important regulatory pathways in stromal cells by influencing miRNA expression. This may alter cellular properties and thus influence stromal cell-based therapeutic applications in regenerative medicine.<\/jats:p>","DOI":"10.3390\/ijms252312589","type":"journal-article","created":{"date-parts":[[2024,11,25]],"date-time":"2024-11-25T03:59:01Z","timestamp":1732507141000},"page":"12589","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Heparin Differentially Regulates the Expression of Specific miRNAs in Mesenchymal Stromal Cells"],"prefix":"10.3390","volume":"25","author":[{"given":"Michaela","family":"Oeller","sequence":"first","affiliation":[{"name":"Department for Transfusion Medicine, University Hospital of Salzburg (SALK), Paracelsus Medical University (PMU) Salzburg, Muellner Hauptstra\u00dfe 48, 5020 Salzburg, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tanja","family":"Schally","sequence":"additional","affiliation":[{"name":"GMP Laboratory, PMU Salzburg, Strubergasse 21, 5020 Salzburg, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Georg","family":"Zimmermann","sequence":"additional","affiliation":[{"name":"Team Biostatistics and Big Medical Data, IDA Lab Salzburg, PMU Salzburg, Strubergasse 16, 5020 Salzburg, Austria"},{"name":"Research Program Biomedical Data Science, PMU Salzburg, Strubergasse 16, 5020 Salzburg, Austria"},{"name":"Department of Artificial Intelligence and Human Interfaces, Faculty of Digital and Analytical Sciences, Paris Lodron University Salzburg, Jakob Haringer Stra\u00dfe 2, 5020 Salzburg, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wanda","family":"Lauth","sequence":"additional","affiliation":[{"name":"Team Biostatistics and Big Medical Data, IDA Lab Salzburg, PMU Salzburg, Strubergasse 16, 5020 Salzburg, Austria"},{"name":"Research Program Biomedical Data Science, PMU Salzburg, Strubergasse 16, 5020 Salzburg, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7204-0058","authenticated-orcid":false,"given":"Katharina","family":"Schallmoser","sequence":"additional","affiliation":[{"name":"Department for Transfusion Medicine, University Hospital of Salzburg (SALK), Paracelsus Medical University (PMU) Salzburg, Muellner Hauptstra\u00dfe 48, 5020 Salzburg, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8692-886X","authenticated-orcid":false,"given":"Eva","family":"Rohde","sequence":"additional","affiliation":[{"name":"Department for Transfusion Medicine, University Hospital of Salzburg (SALK), Paracelsus Medical University (PMU) Salzburg, Muellner Hauptstra\u00dfe 48, 5020 Salzburg, Austria"},{"name":"GMP Laboratory, PMU Salzburg, Strubergasse 21, 5020 Salzburg, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1416-6023","authenticated-orcid":false,"given":"Sandra","family":"Laner-Plamberger","sequence":"additional","affiliation":[{"name":"Department for Transfusion Medicine, University Hospital of Salzburg (SALK), Paracelsus Medical University (PMU) Salzburg, Muellner Hauptstra\u00dfe 48, 5020 Salzburg, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,11,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Rehman, A., Nigam, A., Laino, L., Russo, D., Todisco, C., Esposito, G., Svolacchia, F., Giuzio, F., Desiderio, V., and Ferraro, G. (2023). Mesenchymal Stem Cells in Soft Tissue Regenerative Medicine: A Comprehensive Review. Medicina, 59.","DOI":"10.3390\/medicina59081449"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"136","DOI":"10.1016\/j.reth.2020.01.004","article-title":"Advances in regenerative therapy: A review of the literature and future directions","volume":"14","author":"Ntege","year":"2020","journal-title":"Regen. Ther."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"366","DOI":"10.1186\/s13287-022-03054-0","article-title":"Clinical application of mesenchymal stem cell in regenerative medicine: A narrative review","volume":"13","author":"Margiana","year":"2022","journal-title":"Stem Cell Res. Ther."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Maldonado, V.V., Patel, N.H., Smith, E.E., Barnes, C.L., Gustafson, M.P., Rao, R.R., and Samsonraj, R.M. (2023). Clinical utility of mesenchymal stem\/stromal cells in regenerative medicine and cellular therapy. J. Biol. Eng., 17.","DOI":"10.1186\/s13036-023-00361-9"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Shammaa, R., El-Kadiry, A.E., Abusarah, J., and Rafei, M. (2020). Mesenchymal Stem Cells Beyond Regenerative Medicine. Front. Cell Dev. Biol., 8.","DOI":"10.3389\/fcell.2020.00072"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Pilgrim, C.R., McCahill, K.A., Rops, J.G., Dufour, J.M., Russell, K.A., and Koch, T.G. (2022). A Review of Fetal Bovine Serum in the Culture of Mesenchymal Stromal Cells and Potential Alternatives for Veterinary Medicine. Front. Vet. Sci., 9.","DOI":"10.3389\/fvets.2022.859025"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Oeller, M., Laner-Plamberger, S., Krisch, L., Rohde, E., Strunk, D., and Schallmoser, K. (2021). Human Platelet Lysate for Good Manufacturing Practice-Compliant Cell Production. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22105178"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/j.tibtech.2019.06.002","article-title":"Production and Quality Requirements of Human Platelet Lysate: A Position Statement from the Working Party on Cellular Therapies of the International Society of Blood Transfusion","volume":"38","author":"Schallmoser","year":"2020","journal-title":"Trends Biotechnol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"371","DOI":"10.1016\/j.biomaterials.2015.10.065","article-title":"Human platelet lysate: Replacing fetal bovine serum as a gold standard for human cell propagation?","volume":"76","author":"Burnouf","year":"2016","journal-title":"Biomaterials"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"432","DOI":"10.1186\/s12967-019-02183-0","article-title":"Upregulation of mitotic bookmarking factors during enhanced proliferation of human stromal cells in human platelet lysate","volume":"17","author":"Oeller","year":"2019","journal-title":"J. Transl. Med."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1174","DOI":"10.1016\/j.jcyt.2013.05.006","article-title":"Heparin concentration is critical for cell culture with human platelet lysate","volume":"15","author":"Hemeda","year":"2013","journal-title":"Cytotherapy"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"292","DOI":"10.1016\/j.gene.2015.10.039","article-title":"Effect of heparin on the biological properties and molecular signature of human mesenchymal stem cells","volume":"576","author":"Ling","year":"2016","journal-title":"Gene"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Laner-Plamberger, S., Oeller, M., Poupardin, R., Krisch, L., Hochmann, S., Kalathur, R., Pachler, K., Kreutzer, C., Erdmann, G., and Rohde, E. (2019). Heparin Differentially Impacts Gene Expression of Stromal Cells from Various Tissues. Sci. Rep., 9.","DOI":"10.1038\/s41598-019-43700-x"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"3052","DOI":"10.1093\/ndt\/13.12.3052","article-title":"Effect of heparin on mesangial cell growth and gene expression of matrix proteins","volume":"13","author":"Wang","year":"1998","journal-title":"Nephrol. Dial. Transpl."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1016\/j.bone.2015.04.039","article-title":"Heparin affects human bone marrow stromal cell fate: Promoting osteogenic and reducing adipogenic differentiation and conversion","volume":"78","author":"Simann","year":"2015","journal-title":"Bone"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Laner-Plamberger, S., Oeller, M., Rohde, E., Schallmoser, K., and Strunk, D. (2021). Heparin and Derivatives for Advanced Cell Therapies. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms222112041"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"O\u2019Brien, J., Hayder, H., Zayed, Y., and Peng, C. (2018). Overview of MicroRNA Biogenesis, Mechanisms of Actions, and Circulation. Front. Endocrinol., 9.","DOI":"10.3389\/fendo.2018.00402"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Dexheimer, P.J., and Cochella, L. (2020). MicroRNAs: From Mechanism to Organism. Front. Cell Dev. Biol., 8.","DOI":"10.3389\/fcell.2020.00409"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Palazzo, A.F., and Lee, E.S. (2015). Non-coding RNA: What is functional and what is junk?. Front. Genet., 6.","DOI":"10.3389\/fgene.2015.00002"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Stavast, C.J., and Erkeland, S.J. (2019). The Non-Canonical Aspects of MicroRNAs: Many Roads to Gene Regulation. Cells, 8.","DOI":"10.3390\/cells8111465"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"685","DOI":"10.1002\/bit.28029","article-title":"Biogenesis and mechanisms of microRNA-mediated gene regulation","volume":"119","author":"Rani","year":"2022","journal-title":"Biotechnol. Bioeng."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Catalanotto, C., Cogoni, C., and Zardo, G. (2016). MicroRNA in Control of Gene Expression: An Overview of Nuclear Functions. Int. J. Mol. Sci., 17.","DOI":"10.3390\/ijms17101712"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"441","DOI":"10.1007\/s00018-018-2940-7","article-title":"Regulatory network of miRNA on its target: Coordination between transcriptional and post-transcriptional regulation of gene expression","volume":"76","author":"Pu","year":"2019","journal-title":"Cell Mol. Life Sci."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"92","DOI":"10.1101\/gr.082701.108","article-title":"Most mammalian mRNAs are conserved targets of microRNAs","volume":"19","author":"Friedman","year":"2009","journal-title":"Genome Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"103502","DOI":"10.1016\/j.jri.2022.103502","article-title":"Low molecular weight heparin -induced miRNA changes in peripheral blood mononuclear cells in pregnancies with unexplained recurrent pregnancy loss","volume":"151","author":"Bruno","year":"2022","journal-title":"J. Reprod. Immunol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"26616","DOI":"10.1074\/jbc.M111.224212","article-title":"Heparin impairs angiogenesis through inhibition of microRNA-10b","volume":"286","author":"Shen","year":"2011","journal-title":"J. Biol. Chem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1016\/j.tcb.2014.11.004","article-title":"The biological functions of miRNAs: Lessons from in vivo studies","volume":"25","author":"Vidigal","year":"2015","journal-title":"Trends Cell Biol."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Oduah, E.I., Linhardt, R.J., and Sharfstein, S.T. (2016). Heparin: Past, Present, and Future. Pharmaceuticals, 9.","DOI":"10.3390\/ph9030038"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"354","DOI":"10.1186\/s12967-015-0717-4","article-title":"Mechanical fibrinogen-depletion supports heparin-free mesenchymal stem cell propagation in human platelet lysate","volume":"13","author":"Lener","year":"2015","journal-title":"J. Transl. Med."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"527","DOI":"10.5966\/sctm.2015-0428","article-title":"Heparin Promotes Cardiac Differentiation of Human Pluripotent Stem Cells in Chemically Defined Albumin-Free Medium, Enabling Consistent Manufacture of Cardiomyocytes","volume":"6","author":"Lin","year":"2017","journal-title":"Stem Cells Transl. Med."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Roger, Y., Burmeister, L., Hamm, A., Elger, K., Dittrich-Breiholz, O., Florkemeier, T., and Hoffmann, A. (2020). Heparin Anticoagulant for Human Bone Marrow Does Not Influence In Vitro Performance of Human Mesenchymal Stromal Cells. Cells, 9.","DOI":"10.3390\/cells9071580"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1182\/blood-2014-04-572255","article-title":"Epigenetic and in vivo comparison of diverse MSC sources reveals an endochondral signature for human hematopoietic niche formation","volume":"125","author":"Reinisch","year":"2015","journal-title":"Blood"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1016\/j.ebiom.2019.03.082","article-title":"Pan-cancer analysis on microRNA-associated gene activation","volume":"43","author":"Tan","year":"2019","journal-title":"EBioMedicine"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Xu, P., Wu, Q., Yu, J., Rao, Y., Kou, Z., Fang, G., Shi, X., Liu, W., and Han, H. (2020). A Systematic Way to Infer the Regulation Relations of miRNAs on Target Genes and Critical miRNAs in Cancers. Front. Genet., 11.","DOI":"10.3389\/fgene.2020.00278"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"743","DOI":"10.1016\/S1097-2765(00)00073-3","article-title":"Crystal structure of a ternary FGF-FGFR-heparin complex reveals a dual role for heparin in FGFR binding and dimerization","volume":"6","author":"Schlessinger","year":"2000","journal-title":"Mol. Cell"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1029","DOI":"10.1038\/35039551","article-title":"Crystal structure of fibroblast growth factor receptor ectodomain bound to ligand and heparin","volume":"407","author":"Pellegrini","year":"2000","journal-title":"Nature"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1042\/bj3500245","article-title":"Effect of heparin and liver heparan sulphate on interaction of HepG2-derived transcription factors and their cis-acting elements: Altered potential of hepatocellular carcinoma heparan sulphate","volume":"350","author":"Dudas","year":"2000","journal-title":"Biochem. J."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1608","DOI":"10.1073\/pnas.0707594105","article-title":"MicroRNA-373 induces expression of genes with complementary promoter sequences","volume":"105","author":"Place","year":"2008","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1030","DOI":"10.1038\/nsmb.1841","article-title":"Nuclear-localized tiny RNAs are associated with transcription initiation and splice sites in metazoans","volume":"17","author":"Taft","year":"2010","journal-title":"Nat. Struct. Mol. Biol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1436","DOI":"10.1111\/j.1537-2995.2007.01220.x","article-title":"Human platelet lysate can replace fetal bovine serum for clinical-scale expansion of functional mesenchymal stromal cells","volume":"47","author":"Schallmoser","year":"2007","journal-title":"Transfusion"},{"key":"ref_41","first-page":"1523","article-title":"Preparation of pooled human platelet lysate (pHPL) as an efficient supplement for animal serum-free human stem cell cultures","volume":"32","author":"Schallmoser","year":"2009","journal-title":"J. Vis. Exp."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"349","DOI":"10.1007\/978-1-62703-128-8_22","article-title":"Generation of a pool of human platelet lysate and efficient use in cell culture","volume":"946","author":"Schallmoser","year":"2013","journal-title":"Methods Mol. Biol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1566","DOI":"10.1093\/bioinformatics\/bty852","article-title":"sRNAPrimerDB: Comprehensive primer design and search web service for small non-coding RNAs","volume":"35","author":"Xie","year":"2019","journal-title":"Bioinformatics"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"519","DOI":"10.2144\/000112010","article-title":"Facile means for quantifying microRNA expression by real-time PCR","volume":"39","author":"Shi","year":"2005","journal-title":"Biotechniques"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"192","DOI":"10.2144\/btn-2019-0065","article-title":"qPCR-based methods for expression analysis of miRNAs","volume":"67","author":"Forero","year":"2019","journal-title":"Biotechniques"},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Kang, K., Zhang, X., Liu, H., Wang, Z., Zhong, J., Huang, Z., Peng, X., Zeng, Y., Wang, Y., and Yang, Y. (2012). A novel real-time PCR assay of microRNAs using S-Poly(T), a specific oligo(dT) reverse transcription primer with excellent sensitivity and specificity. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0048536"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"588","DOI":"10.1016\/j.ygyno.2013.06.026","article-title":"Selection and validation of endogenous controls for microRNA expression studies in endometrioid endometrial cancer tissues","volume":"130","author":"Torres","year":"2013","journal-title":"Gynecol. Oncol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1333","DOI":"10.1373\/clinchem.2015.239459","article-title":"Data Normalization Strategies for MicroRNA Quantification","volume":"61","author":"Schwarzenbach","year":"2015","journal-title":"Clin. Chem."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"5529","DOI":"10.1038\/sj.onc.1205748","article-title":"Human GLI2 and GLI1 are part of a positive feedback mechanism in Basal Cell Carcinoma","volume":"21","author":"Regl","year":"2002","journal-title":"Oncogene"},{"key":"ref_50","unstructured":"R-Core-Team (2024). A Language and Environment for Statistical Computing, R Foundation for Statistical Computing. Available online: https:\/\/www.R-project.org\/."},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Wickham, H. (2016). ggplot2: Elegant Graphics for Data Analysis, Springer.","DOI":"10.1007\/978-3-319-24277-4_9"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"W566","DOI":"10.1093\/nar\/gkv468","article-title":"ClustVis: A web tool for visualizing clustering of multivariate data using Principal Component Analysis and heatmap","volume":"43","author":"Metsalu","year":"2015","journal-title":"Nucleic Acids Res."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/23\/12589\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T16:38:00Z","timestamp":1760114280000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/23\/12589"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,11,23]]},"references-count":52,"journal-issue":{"issue":"23","published-online":{"date-parts":[[2024,12]]}},"alternative-id":["ijms252312589"],"URL":"https:\/\/doi.org\/10.3390\/ijms252312589","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,11,23]]}}}