{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T19:46:18Z","timestamp":1785267978508,"version":"3.55.0"},"reference-count":39,"publisher":"Oxford University Press (OUP)","issue":"1","license":[{"start":{"date-parts":[[2024,10,29]],"date-time":"2024-10-29T00:00:00Z","timestamp":1730160000000},"content-version":"vor","delay-in-days":302,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"name":"Centre or Education Services, Ministry of Education, Culture, Research"},{"name":"Technology of the Republic of Indonesia"},{"name":"Indonesia Endowment Funds for Education"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2024,1,11]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>This protocol introduces a streamlined and efficient method for isolating human fibroblast from skin primary cell culture with a specific focus on its application to keloid, hypertrophic scar, and normal skin biopsies. Additionally, the absence of suitable animal models for keloid and hypertrophic scar has led preclinical research to rely on in vitro studies using primary cell cultures. This approach addresses the challenges of existing protocols in terms of time, cost, equipment, and technical expertise required. The method involves derivation, culture, and characterization analysis including cell proliferation, migration, and fibroblastic marker (Vimentin, CD90, CD73, and CD105) expression. Our study yielded high amounts of fibroblast from tested skin explants while maintaining their in vivo-like characteristics and behaviour. Immunostaining assay confirmed that the cultivated fibroblast was positively expressed Vimentin. Flowcytometry results showed high expression of CD90 and CD73 while relatively showing lower expression of CD105. Fibroblast derived from keloid tissue showed the highest rate of proliferation and migration ability compared to the other samples. These findings suggest an efficient and reproducible technique to cultivate high qualified fibroblast from human skin in normal or pathological condition, particularly for keloid and hypertrophic scar. The application of this protocol provides a foundation for further studies to investigate the progression and potential intervention of aberrant fibrotic dermatological disorder, in vitro.<\/jats:p>","DOI":"10.1093\/biomethods\/bpae082","type":"journal-article","created":{"date-parts":[[2024,10,29]],"date-time":"2024-10-29T21:54:23Z","timestamp":1730238863000},"source":"Crossref","is-referenced-by-count":8,"title":["Efficient protocol for isolating human fibroblast from primary skin cell cultures: application to keloid, hypertrophic scar, and normal skin biopsies"],"prefix":"10.1093","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1782-7509","authenticated-orcid":false,"given":"Sri Suciati","family":"Ningsih","sequence":"first","affiliation":[{"name":"Doctoral Program in Biomedical Sciences, Faculty of Medicine, Universitas Indonesia , Jakarta 10430,","place":["Indonesia"]},{"name":"Faculty of Medicine, Universitas Muhammadiyah Prof Dr Hamka , Jakarta 12130,","place":["Indonesia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0471-5583","authenticated-orcid":false,"given":"Sri Widia A","family":"Jusman","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, Faculty of Medicine, Universitas Indonesia , Jakarta 10430,","place":["Indonesia"]},{"name":"Center of Hypoxia and Oxidative Stress Studies, Faculty of Medicine, Universitas Indonesia , Jakarta 10430,","place":["Indonesia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0553-7109","authenticated-orcid":false,"given":"Rahimi","family":"Syaidah","sequence":"additional","affiliation":[{"name":"Doctoral Program in Biomedical Sciences, Faculty of Medicine, Universitas Indonesia , Jakarta 10430,","place":["Indonesia"]},{"name":"Department of Histology, Faculty of Medicine, Universitas Indonesia , Jakarta 10430,","place":["Indonesia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-5390-4551","authenticated-orcid":false,"given":"Raisa","family":"Nauli","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, Faculty of Medicine, Universitas Indonesia , Jakarta 10430,","place":["Indonesia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8120-3138","authenticated-orcid":false,"given":"Fadilah","family":"Fadilah","sequence":"additional","affiliation":[{"name":"Doctoral Program in Biomedical Sciences, Faculty of Medicine, Universitas Indonesia , Jakarta 10430,","place":["Indonesia"]},{"name":"Department of Medical Chemistry, Faculty of Medicine, Universitas Indonesia , Jakarta 10430,","place":["Indonesia"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2024,10,29]]},"reference":[{"key":"2024121101594561000_bpae082-B1","doi-asserted-by":"publisher","first-page":"3852","DOI":"10.1016\/j.cell.2021.06.024","article-title":"Fibroblasts: origins, definitions, and functions in health and disease","volume":"184","author":"Plikus","year":"2021","journal-title":"Cell"},{"key":"2024121101594561000_bpae082-B2","doi-asserted-by":"publisher","first-page":"37","DOI":"10.1016\/j.matbio.2016.01.013","article-title":"Keloids: the paradigm of skin fibrosis\u2014Pathomechanisms and treatment","volume":"51","author":"Andrews","year":"2016","journal-title":"Matrix Biol"},{"key":"2024121101594561000_bpae082-B3","doi-asserted-by":"publisher","first-page":"S3","DOI":"10.1097\/DSS.0000000000000819","article-title":"Keloids and hypertrophic scars: pathophysiology, classification, and treatment","volume":"43","author":"Berman","year":"2017","journal-title":"Dermatol Surg"},{"key":"2024121101594561000_bpae082-B4","doi-asserted-by":"publisher","first-page":"606","DOI":"10.3390\/ijms18030606","article-title":"Keloid and hypertrophic scars are the result of chronic inflammation in the reticular dermis","volume":"18","author":"Ogawa","year":"2017","journal-title":"Int J Mol Sci"},{"key":"2024121101594561000_bpae082-B5","doi-asserted-by":"publisher","first-page":"711","DOI":"10.3390\/ijms19030711","article-title":"Recent understandings of biology, prophylaxis and treatment strategies for hypertrophic scars and keloids","volume":"19","author":"Lee","year":"2018","journal-title":"Int J Mol Sci"},{"key":"2024121101594561000_bpae082-B6","doi-asserted-by":"publisher","first-page":"77","DOI":"10.1089\/wound.2018.0828","article-title":"Animal Models for Studies of Keloid Scarring","volume":"8","author":"Supp","year":"2019","journal-title":"Adv Wound Care (New Rochelle)"},{"key":"2024121101594561000_bpae082-B7","doi-asserted-by":"publisher","first-page":"385","DOI":"10.1177\/2051415818761526","article-title":"Human foreskin fibroblasts: from waste bag to important biomedical applications","volume":"11","author":"Oliveira","year":"2018","journal-title":"J Clin Urol"},{"key":"2024121101594561000_bpae082-B8","doi-asserted-by":"publisher","first-page":"89","DOI":"10.1007\/s00423-015-1343-6","article-title":"Isolation and culture of primary human pancreatic stellate cells that reflect the context of their tissue of origin","volume":"401","author":"Strobel","year":"2016","journal-title":"Langenbecks Arch Surg"},{"key":"2024121101594561000_bpae082-B9","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1111\/cpr.12334","article-title":"Explant culture: an advantageous method for isolation of mesenchymal stem cells from human tissues","volume":"50","author":"Hendijani","year":"2017","journal-title":"Cell Prolif"},{"key":"2024121101594561000_bpae082-B10","doi-asserted-by":"publisher","first-page":"40","DOI":"10.1038\/s41419-020-03323-x","article-title":"HOXA5 counteracts the function of pathological scar-derived fibroblasts by partially activating p53 signaling","volume":"12","author":"Liang","year":"2021","journal-title":"Cell Death Dis"},{"key":"2024121101594561000_bpae082-B11","doi-asserted-by":"publisher","first-page":"23","DOI":"10.21037\/sci.2019.08.05","article-title":"Isolating human dermal fibroblasts using serial explant culture","volume":"6","author":"Nejaddehbashi","year":"2019","journal-title":"Stem Cell Investig"},{"key":"2024121101594561000_bpae082-B12","doi-asserted-by":"publisher","first-page":"566607","DOI":"10.3389\/fimmu.2020.566607","article-title":"A Method for Isolating and Culturing Skin Cells: application to Endothelial Cells, Fibroblasts, Keratinocytes, and Melanocytes From Punch Biopsies in Systemic Sclerosis Skin","volume":"11","author":"Henrot","year":"2020","journal-title":"Front Immunol"},{"key":"2024121101594561000_bpae082-B13","first-page":"245","volume-title":"Biochem Biophys Res Commun","author":"Raktoe","year":"2021"},{"key":"2024121101594561000_bpae082-B14","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1088\/1758-5090\/ad2b06","article-title":"3D modeling of normal skin and cutaneous squamous cell carcinoma. A comparative study in 2D cultures, spheroids, and 3D bioprinted systems","volume":"16","author":"Kurzyk","year":"2024","journal-title":"Biofabrication"},{"key":"2024121101594561000_bpae082-B15","doi-asserted-by":"publisher","first-page":"869069","DOI":"10.3389\/fcell.2022.869069","article-title":"Impact of vimentin on regulation of cell signaling and matrix remodeling","volume":"10","author":"Ostrowska-Podhorodecka","year":"2022","journal-title":"Front Cell Dev Biol"},{"key":"2024121101594561000_bpae082-B16","doi-asserted-by":"publisher","first-page":"929495","DOI":"10.3389\/fcell.2022.929495","article-title":"Expression of vimentin alters cell mechanics, cell-cell adhesion, and gene expression profiles suggesting the induction of a hybrid EMT in human mammary epithelial cells","volume":"10","author":"Sivagurunathan","year":"2022","journal-title":"Front Cell Dev Biol"},{"key":"2024121101594561000_bpae082-B17","doi-asserted-by":"publisher","first-page":"85","DOI":"10.1159\/000445096","article-title":"Fibroblasts and mesenchymal stromal\/stem cells are phenotypically indistinguishable","volume":"136","author":"Denu","year":"2016","journal-title":"Acta Haematol"},{"key":"2024121101594561000_bpae082-B18","doi-asserted-by":"publisher","first-page":"441","DOI":"10.1242\/dmm.044164","article-title":"Understanding the impact of fibroblast heterogeneity on skin fibrosis","volume":"13","author":"Griffin","year":"2020","journal-title":"Dis Model Mech"},{"key":"2024121101594561000_bpae082-B19","doi-asserted-by":"publisher","first-page":"1403","DOI":"10.1248\/bpb.b21-00058","article-title":"Establishment of three types of immortalized human skin stem cell lines derived from the single donor","volume":"44","author":"Inoue","year":"2021","journal-title":"Biol Pharm Bull"},{"key":"2024121101594561000_bpae082-B20","doi-asserted-by":"publisher","first-page":"8430","DOI":"10.7150\/THNO.59661","article-title":"Self-assembly of differentiated progenitor cells facilitates spheroid human skin organoid formation and planar skin regeneration","volume":"11","author":"Ebner-Peking","year":"2021","journal-title":"Theranostics"},{"key":"2024121101594561000_bpae082-B21","doi-asserted-by":"publisher","first-page":"101378","DOI":"10.1016\/j.xpro.2022.101378","article-title":"Isolation of five different primary cell types from a single sample of human skin","volume":"3","author":"Kabacik","year":"2022","journal-title":"STAR Protoc"},{"key":"2024121101594561000_bpae082-B22","doi-asserted-by":"publisher","first-page":"73","DOI":"10.1007\/7651_2020_332","article-title":"Isolation, culture, cryopreservation, and preparation of umbilical cord-derived mesenchymal stem cells as a final cellular product under good manufacturing practices\u2013compliant conditions","volume":"2286","author":"Aydo\u011fdu","year":"2021","journal-title":"Methods Mol Biol"},{"key":"2024121101594561000_bpae082-B23","doi-asserted-by":"publisher","first-page":"e714","DOI":"10.1002\/cpz1.714","article-title":"Simple, fast, and efficient method for derivation of dermal fibroblasts from skin biopsies","volume":"3","author":"Iannello","year":"2023","journal-title":"Curr Protoc"},{"key":"2024121101594561000_bpae082-B24","doi-asserted-by":"publisher","first-page":"e0142398","DOI":"10.1371\/journal.pone.0142398","article-title":"The performance of the four anaerobic blood culture bottles BacT\/ALERT-FN,-FN Plus, bactec-plus and-lytic in detection of anaerobic bacteria and identification by direct MALDI-TOF MS","volume":"10","author":"Almuhayawi","year":"2015","journal-title":"PLoS One"},{"key":"2024121101594561000_bpae082-B25","doi-asserted-by":"publisher","first-page":"285","DOI":"10.3389\/fcimb.2019.00285","article-title":"Comparison of the performance of three blood culture systems in a chinese tertiary-care hospital","volume":"9","author":"Li","year":"2019","journal-title":"Front Cell Infect Microbiol"},{"key":"2024121101594561000_bpae082-B26","doi-asserted-by":"publisher","first-page":"101172","DOI":"10.1016\/j.xpro.2022.101172","article-title":"Isolation and feeder-free primary culture of four cell types from a single human skin sample","volume":"3","author":"Supp","year":"2022","journal-title":"STAR Protoc"},{"key":"2024121101594561000_bpae082-B27","doi-asserted-by":"publisher","first-page":"105","DOI":"10.1186\/s13578-022-00842-2","article-title":"Human skin dermis-derived fibroblasts are a kind of functional mesenchymal stromal cells: judgements from surface markers, biological characteristics, to therapeutic efficacy","volume":"12","author":"Tai","year":"2022","journal-title":"Cell Biosci"},{"key":"2024121101594561000_bpae082-B28","doi-asserted-by":"publisher","first-page":"368","DOI":"10.3390\/cells9020368","article-title":"Fibroblasts from the human skin dermo-hypodermal junction are distinct from dermal papillary and reticular fibroblasts and from mesenchymal stem cells and exhibit a specific molecular profile related to extracellular matrix organization and modeling","volume-title":"Cells","author":"Haydont","year":"2020"},{"key":"2024121101594561000_bpae082-B29","doi-asserted-by":"publisher","first-page":"5327","DOI":"10.1172\/JCI130239","article-title":"Dermal adipose tissue has high plasticity and undergoes reversible dedifferentiation in mice","volume":"129","author":"Zhang","year":"2019","journal-title":"J Clin Invest"},{"key":"2024121101594561000_bpae082-B30","doi-asserted-by":"publisher","first-page":"75114","DOI":"10.18632\/oncotarget.20540","article-title":"Dietary menthol-induced TRPM8 activation enhances WAT 1C \u201cbrowning 1D\u201d and ameliorates diet-induced obesity","volume":"8","author":"Jiang","year":"2017","journal-title":"Oncotarget"},{"key":"2024121101594561000_bpae082-B31","doi-asserted-by":"publisher","first-page":"24389","DOI":"10.1038\/srep24389","article-title":"Epithelial cell migration requires the interaction between the vimentin and keratin intermediate filaments","volume":"6","author":"Velez-Delvalle","year":"2016","journal-title":"Sci Rep"},{"key":"2024121101594561000_bpae082-B32","doi-asserted-by":"publisher","first-page":"1487","DOI":"10.5114\/aoms\/131088","article-title":"Flow cytometric characterization of cell surface markers to differentiate between fibroblasts and mesenchymal stem cells of different origin","volume":"19","author":"Sober","year":"2023","journal-title":"Arch Med Sci"},{"key":"2024121101594561000_bpae082-B33","first-page":"38","author":"Nauli","year":"2022"},{"key":"2024121101594561000_bpae082-B34","doi-asserted-by":"publisher","first-page":"422","DOI":"10.1111\/wrr.12793","article-title":"Keloid scarring or disease: unresolved quasi-neoplastic tendencies in the human skin","volume":"28","author":"Ud-Din","year":"2020","journal-title":"Wound Repair Regen"},{"key":"2024121101594561000_bpae082-B35","doi-asserted-by":"publisher","first-page":"3709","DOI":"10.1038\/s41467-021-24110-y","article-title":"Single-cell RNA-seq reveals fibroblast heterogeneity and increased mesenchymal fibroblasts in human fibrotic skin diseases","volume":"12","author":"Deng","year":"2021","journal-title":"Nat Commun"},{"key":"2024121101594561000_bpae082-B36","doi-asserted-by":"publisher","first-page":"e75600","DOI":"10.1371\/journal.pone.0075600","article-title":"Site-specific keloid fibroblasts alter the behaviour of normal skin and normal scar fibroblasts through paracrine signalling","volume":"8","author":"Ashcroft","year":"2013","journal-title":"PLoS One"},{"key":"2024121101594561000_bpae082-B37","doi-asserted-by":"publisher","first-page":"e0253669","DOI":"10.1371\/journal.pone.0253669","article-title":"Therapeutic candidates for keloid scars identified by qualitative review of scratch assay research for wound healing","volume-title":"PLoS One","author":"Alishahedani","year":"2021"},{"key":"2024121101594561000_bpae082-B38","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/ijms22042198","article-title":"Ameliorating fibrotic phenotypes of keloid dermal fibroblasts through an epidermal growth factor-mediated extracellular matrix remodeling","volume":"22","author":"Kim","year":"2021","journal-title":"Int J Mol Sci"},{"key":"2024121101594561000_bpae082-B39","doi-asserted-by":"publisher","first-page":"95","DOI":"10.1016\/j.jdermsci.2016.03.003","article-title":"Bone marrow derived mesenchymal stem cells inhibit the proliferative and profibrotic phenotype of hypertrophic scar fibroblasts and keloid fibroblasts through paracrine signaling","volume":"83","author":"Fang","year":"2016","journal-title":"J Dermatol Sci"}],"container-title":["Biology Methods and Protocols"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/biomethods\/advance-article-pdf\/doi\/10.1093\/biomethods\/bpae082\/60212554\/bpae082.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/biomethods\/article-pdf\/9\/1\/bpae082\/61019471\/bpae082.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/biomethods\/article-pdf\/9\/1\/bpae082\/61019471\/bpae082.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,12,11]],"date-time":"2024-12-11T03:09:19Z","timestamp":1733886559000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/biomethods\/article\/doi\/10.1093\/biomethods\/bpae082\/7849646"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024]]},"references-count":39,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2024,1,11]]}},"URL":"https:\/\/doi.org\/10.1093\/biomethods\/bpae082","relation":{},"ISSN":["2396-8923"],"issn-type":[{"value":"2396-8923","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2024]]},"published":{"date-parts":[[2024]]},"article-number":"bpae082"}}