{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T22:50:57Z","timestamp":1792191057364,"version":"4.4.0"},"reference-count":46,"publisher":"MDPI AG","issue":"13","license":[{"start":{"date-parts":[[2022,6,26]],"date-time":"2022-06-26T00:00:00Z","timestamp":1656201600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Design-based stereology is the gold standard for obtaining unbiased quantitative morphological data on volume, surface area, and length, as well as the number of tissues, cells or organelles. In cardiac research, the introduction of a stereological method to unbiasedly estimate the number of cardiomyocytes has considerably increased the use of stereology. Since its original description, various modifications to this method have been described. A particular field in which this method has been employed is the normal developmental life cycle of cardiomyocytes after birth, and particularly the question of when, during postnatal development, cardiomyocytes lose their capacity to divide and proliferate, and thus their inherent regenerative ability. This field is directly related to a second major application of stereology in recent years, addressing the question of what consequences intrauterine growth restriction has on the development of the heart, particularly of cardiomyocytes. Advances have also been made regarding the quantification of nerve fibers and collagen deposition as measures of heart innervation and fibrosis. In the present review article, we highlight the methodological progress made in the last 20 years and demonstrate how stereology has helped to gain insight into the process of normal cardiac development, and how it is affected by intrauterine growth restriction.<\/jats:p>","DOI":"10.3390\/cells11132032","type":"journal-article","created":{"date-parts":[[2022,6,26]],"date-time":"2022-06-26T09:00:13Z","timestamp":1656234013000},"page":"2032","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Methodological Progress of Stereology in Cardiac Research and Its Application to Normal and Pathological Heart Development"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7827-4867","authenticated-orcid":false,"given":"Christian","family":"M\u00fchlfeld","sequence":"first","affiliation":[{"name":"Institute of Functional and Applied Anatomy, Hannover Medical School, 30625 Hannover, Germany"},{"name":"Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), Member of the German Center for Lung Research (DZL), 30625 Hannover, Germany"},{"name":"Research Core Unit Electron Microscopy, Hannover Medical School, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0022-6109","authenticated-orcid":false,"given":"Julia","family":"Schipke","sequence":"additional","affiliation":[{"name":"Institute of Functional and Applied Anatomy, Hannover Medical School, 30625 Hannover, Germany"},{"name":"Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), Member of the German Center for Lung Research (DZL), 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,6,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1053\/plac.2002.0878","article-title":"The \u2018reference trap\u2019 revisited: Examples of the dangers in using ratios to describe fetoplacental angiogenesis and trophoblast turnover","volume":"24","author":"Mayhew","year":"2003","journal-title":"Placenta"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1100","DOI":"10.1681\/ASN.V1051100","article-title":"Stereological methods and their application in kidney research","volume":"10","author":"Nyengaard","year":"1999","journal-title":"J. Am. Soc. Nephrol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"394","DOI":"10.1164\/rccm.200809-1522ST","article-title":"An official research policy statement of the American Thoracic Society\/European Respiratory Society: Standards for quantitative assessment of lung structure","volume":"181","author":"Hsia","year":"2010","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"L15","DOI":"10.1152\/ajplung.00429.2012","article-title":"Quantitative microscopy of the lung: A problem-based approach. Part 1: Basic principles of lung stereology","volume":"305","author":"Ochs","year":"2013","journal-title":"Am. J. Physiol. Lung Cell Mol. Physiol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"L205","DOI":"10.1152\/ajplung.00429.2012","article-title":"Quantitative microscopy of the lung: A problem-based approach. Part 2: Stereological parameters and study designs in various diseases of the respiratory tract","volume":"305","author":"Ochs","year":"2013","journal-title":"Am. J. Physiol. Lung Cell Mol. Physiol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1482","DOI":"10.4067\/S0717-95022017000401482","article-title":"Tips for studies with quantitative morphology (morphometry and stereology)","volume":"35","year":"2017","journal-title":"Int. J. Morphol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1016\/j.carpath.2008.10.015","article-title":"A review of state-of-the-art stereology for better quantitative 3D morphology in cardiac research","volume":"19","author":"Nyengaard","year":"2010","journal-title":"Cardiovasc. Pathol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1016\/j.aanat.2012.10.010","article-title":"Quantitative morphology of the vascularisation of organs: A stereological approach illustrated using the cardiac circulation","volume":"196","year":"2014","journal-title":"Ann. Anat."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1111\/j.1365-2818.1984.tb02501.x","article-title":"The unbiased estimation of number and sizes of arbitrary particles using the disector","volume":"134","author":"Sterio","year":"1984","journal-title":"J. Microsc."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1007\/s00395-005-0524-9","article-title":"Design-based stereological estimation of the total number of cardiac myocytes in histological sections","volume":"100","author":"Nyengaard","year":"2005","journal-title":"Basic Res. Cardiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"539","DOI":"10.1111\/joa.12236","article-title":"The number of cardiac myocytes in the hypertrophic and hypotrophic left ventricle of the obese and calorie-restricted mouse heart","volume":"225","author":"Schipke","year":"2014","journal-title":"J. Anat."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"796","DOI":"10.1203\/01.PDR.0000157726.65492.CD","article-title":"Effect of intrauterine growth restriction on the number of cardiomyocytes in rat hearts","volume":"57","author":"Corstius","year":"2005","journal-title":"Pediatr. Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/j.ymeth.2020.06.002","article-title":"Stereological estimation of cardiomyocyte number and proliferation","volume":"190","author":"Silva","year":"2021","journal-title":"Methods"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"4790","DOI":"10.7150\/thno.57456","article-title":"Thyroid hormone plus dual-specificity phosphatase-5 siRNA increases the number of cardiac muscle cells and improves left ventricular contractile function in chronic doxorubicin-injured hearts","volume":"11","author":"Tan","year":"2021","journal-title":"Theranostics"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"390","DOI":"10.1164\/rccm.201709-1838LE","article-title":"Flow Cytometry Underestimates and Planimetry Overestimates Alveolar Epithelial Type 2 Cell Expansion after Lung Injury","volume":"198","author":"Jansing","year":"2018","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1111\/jmi.12800","article-title":"Estimation of absolute number of alveolar epithelial type 2 cells in mouse lungs: A comparison between stereology and flow cytometry","volume":"275","author":"Dzhuraev","year":"2019","journal-title":"J. Microsc."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1402","DOI":"10.1152\/japplphysiol.01013.2009","article-title":"An unbiased stereological method for efficiently quantifying the innervation of the heart and other organs based on total length estimations","volume":"108","author":"Papadakis","year":"2010","journal-title":"J. Appl. Physiol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"518","DOI":"10.1111\/joa.12151","article-title":"Allometry of left ventricular myocardial innervation","volume":"224","author":"Schipke","year":"2014","journal-title":"J. Anat."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1007\/s13402-011-0035-7","article-title":"Rapid quantification of myocardial fibrosis: A new macro-based automated analysis","volume":"34","author":"Hadi","year":"2011","journal-title":"Cell. Oncol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"114","DOI":"10.1186\/1746-1596-9-114","article-title":"Quantification of myocardial fibrosis by digital image analysis and interactive stereology","volume":"9","author":"Daunoravicius","year":"2014","journal-title":"Diagn. Pathol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1019","DOI":"10.1152\/japplphysiol.00987.2016","article-title":"Assessment of cardiac fibrosis: A morphometric method comparison for collagen quantification","volume":"122","author":"Schipke","year":"2017","journal-title":"J. Appl. Physiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"L341","DOI":"10.1152\/ajplung.00329.2013","article-title":"Alterations of mouse lung tissue dimensions during processing for morphometry: A comparison of methods","volume":"306","author":"Schneider","year":"2014","journal-title":"Am. J. Physiol. Lung Cell Mol. Physiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1446","DOI":"10.1073\/pnas.1214608110","article-title":"Cardiomyocyte proliferation contributes to heart growth in young humans","volume":"110","author":"Mollova","year":"2013","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1566","DOI":"10.1016\/j.cell.2015.05.026","article-title":"Dynamics of cell generation and turnover in the human heart","volume":"161","author":"Bergmann","year":"2015","journal-title":"Cell"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1026","DOI":"10.1016\/j.cell.2015.10.035","article-title":"No evidence for cardiomyocyte number expansion in preadolescent mice","volume":"163","author":"Alkass","year":"2015","journal-title":"Cell"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"565","DOI":"10.1016\/j.cell.2014.03.032","article-title":"The oxygen-rich postnatal environment induces cardiomyocyte cell-cycle arrest through DNA damage response","volume":"157","author":"Puente","year":"2014","journal-title":"Cell"},{"key":"ref_27","first-page":"H2183","article-title":"Cardiomyocyte DNA synthesis and binucleation during murine development","volume":"271","author":"Soonpaa","year":"1996","journal-title":"Am. J. Physiol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"365","DOI":"10.1093\/cvr\/cvq005","article-title":"Cardiomyocyte cell cycle control and growth estimation in vivo\u2014An analysis based on cardiomyocyte nuclei","volume":"86","author":"Walsh","year":"2010","journal-title":"Cardiovasc. Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"795","DOI":"10.1016\/j.cell.2014.03.035","article-title":"A proliferative burst during preadolescence establishes the final cardiomyocyte number","volume":"157","author":"Naqvi","year":"2014","journal-title":"Cell"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1113\/JP275339","article-title":"Sustained cardiac programming by short-term juvenile exercise training in male rats","volume":"596","author":"Asif","year":"2018","journal-title":"J. Physiol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"400","DOI":"10.1016\/j.cardiores.2007.06.026","article-title":"Growth hormone increases the proliferation of existing cardiac myocytes and the total number of cardiac myocytes in the rat heart","volume":"76","author":"Christoffersen","year":"2007","journal-title":"Cardiovasc. Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1007\/s00395-007-0684-x","article-title":"Effect of voluntary exercise on number and volume of cardiomyocytes and their mitochondria in the mouse left ventricle","volume":"103","author":"Eisele","year":"2008","journal-title":"Basic Res. Cardiol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"552","DOI":"10.1161\/CIRCRESAHA.119.315408","article-title":"Polyploidy in Cardiomyocytes: Roadblock to Heart Regeneration?","volume":"126","author":"Derks","year":"2020","journal-title":"Circ. Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1346","DOI":"10.1038\/ng.3929","article-title":"Frequency of mononuclear diploid cardiomyocytes underlies natural variation in heart regeneration","volume":"49","author":"Patterson","year":"2017","journal-title":"Nat. Genet."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"322","DOI":"10.1016\/j.ajpath.2013.06.035","article-title":"Polyploidization in liver tissue","volume":"184","author":"Gentric","year":"2014","journal-title":"Am. J. Pathol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1166","DOI":"10.1016\/j.cub.2012.05.016","article-title":"Hypertrophy and unconventional cell division of hepatocytes underlie liver regeneration","volume":"22","author":"Miyaoka","year":"2012","journal-title":"Curr. Biol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"3105","DOI":"10.1016\/j.celrep.2020.02.034","article-title":"Direct Comparison of Mononucleated and Binucleated Cardiomyocytes Reveals Molecular Mechanisms Underlying Distinct Proliferative Competencies","volume":"30","author":"Windmueller","year":"2020","journal-title":"Cell Rep."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"168","DOI":"10.1016\/j.reprotox.2020.10.005","article-title":"Intrauterine growth restriction: Clinical consequences on health and disease at adulthood","volume":"99","author":"Armengaud","year":"2021","journal-title":"Reprod. Toxicol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"564","DOI":"10.1136\/bmj.298.6673.564","article-title":"Growth in utero, blood pressure in childhood and adult life, and mortality from cardiovascular disease","volume":"298","author":"Barker","year":"1989","journal-title":"BMJ"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"819","DOI":"10.1093\/eurjhf\/hfq069","article-title":"The early origins of chronic heart failure: Impaired placental growth and initiation of insulin resistance in childhood","volume":"12","author":"Barker","year":"2010","journal-title":"Eur. J. Heart Fail"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"R48","DOI":"10.1152\/ajpregu.00180.2017","article-title":"IUGR decreases cardiomyocyte endowment and alters cardiac metabolism in a sex- and cause-of-IUGR-specific manner","volume":"315","author":"Botting","year":"2018","journal-title":"Am. J. Physiol. Regul. Integr. Comp. Physiol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1002\/ar.21084","article-title":"Effect of maternal protein restriction during pregnancy and lactation on the number of cardiomyocytes in the postproliferative weanling rat heart","volume":"293","author":"Lim","year":"2010","journal-title":"Anat. Rec."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"e000531","DOI":"10.1161\/JAHA.113.000531","article-title":"Chronic hypoxemia in late gestation decreases cardiomyocyte number but does not change expression of hypoxia-responsive genes","volume":"3","author":"Botting","year":"2014","journal-title":"J. Am. Heart Assoc."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"557","DOI":"10.1007\/s00418-017-1587-z","article-title":"Experimentally induced intrauterine growth restriction in rabbits leads to differential remodelling of left versus right ventricular myocardial microstructure","volume":"148","author":"Schipke","year":"2017","journal-title":"Histochem. Cell Biol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"618","DOI":"10.1017\/S2040174417000381","article-title":"Small size at birth predicts decreased cardiomyocyte number in the adult bovine heart","volume":"8","author":"Vranas","year":"2017","journal-title":"J. Dev. Orig. Health Dis."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1111\/j.1365-2818.1986.tb02764.x","article-title":"Stereology of arbitrary particles. A review of unbiased number and size estimators and the presentation of some new ones, in memory of William, R. Thompson","volume":"143","author":"Gundersen","year":"1986","journal-title":"J. Microsc."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/11\/13\/2032\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:38:41Z","timestamp":1760139521000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/11\/13\/2032"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,6,26]]},"references-count":46,"journal-issue":{"issue":"13","published-online":{"date-parts":[[2022,7]]}},"alternative-id":["cells11132032"],"URL":"https:\/\/doi.org\/10.3390\/cells11132032","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,6,26]]}}}