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Characterization of keratinocytes, fibroblasts and melanocytes isolated from human skin using gene markers

Cells isolated from skin have wide applications in studies of the pathogenesis of skin-related diseases and the construction of 3D skin equivalents. This study aimed to isolate keratinocytes, fibroblasts, and melanocytes from human foreskin and characterize the purity of the cell types. Keratinocyte...

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Main Authors:	Wong, Pau-Ring, Nur Fatin Nabilah Mohd Sahardi,, Tan, Jen-Kit, Chua, Kien-Hui, Wan Zurinah Wan Ngah,, Suzana Makpol,
Format:	Article
Language:	English
Published:	Penerbit Universiti Kebangsaan Malaysia 2022
Online Access:	http://journalarticle.ukm.my/19477/1/13.pdf http://journalarticle.ukm.my/19477/ https://www.ukm.my/jsm/malay_journals/jilid51bil5_2022/KandunganJilid51Bil5_2022.html
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spelling	my-ukm.journal.194772022-08-26T01:43:49Z http://journalarticle.ukm.my/19477/ Characterization of keratinocytes, fibroblasts and melanocytes isolated from human skin using gene markers Wong, Pau-Ring Nur Fatin Nabilah Mohd Sahardi, Tan, Jen-Kit Chua, Kien-Hui Wan Zurinah Wan Ngah, Suzana Makpol, Cells isolated from skin have wide applications in studies of the pathogenesis of skin-related diseases and the construction of 3D skin equivalents. This study aimed to isolate keratinocytes, fibroblasts, and melanocytes from human foreskin and characterize the purity of the cell types. Keratinocytes, fibroblasts, and melanocytes from human foreskin were isolated by differential trypsinization and media selection. The purity of the cell types was characterized based on the expression of gene markers. The assessment of gene marker expression involved RNA extraction, primer design, quantitative polymerase chain reaction (qPCR) and immunocytochemical staining. Our results showed that in cocultures of keratinocytes and fibroblasts isolated from the dermis, fibroblasts could be separated from keratinocytes by quick trypsinization and culture in Dulbecco’s modified Eagle’s medium. The remaining keratinocytes are cultured in Epilife medium. Melanocytes in cocultures of melanocytes and keratinocytes isolated from the epidermis could be selected by changing Epilife medium to M254 medium. Gene marker results suggested that cytokeratin 14 (CK14) is a suitable marker for keratinocytes, elastin (ELN) is a suitable marker for fibroblasts, and tyrosinase (TYR) and tyrosinase-related proteins 1 and 2 (TYRP1 and TYRP2) are suitable markers for melanocytes. In conclusion, keratinocytes, fibroblasts, and melanocytes can be isolated from the same human foreskin sample by differential trypsinization and media selection techniques and characterized by suitable gene markers. This finding will aid in the isolation of pure skin cell types for various applications in regenerative medicine and toxicity studies. Penerbit Universiti Kebangsaan Malaysia 2022-05 Article PeerReviewed application/pdf en http://journalarticle.ukm.my/19477/1/13.pdf Wong, Pau-Ring and Nur Fatin Nabilah Mohd Sahardi, and Tan, Jen-Kit and Chua, Kien-Hui and Wan Zurinah Wan Ngah, and Suzana Makpol, (2022) Characterization of keratinocytes, fibroblasts and melanocytes isolated from human skin using gene markers. Sains Malaysiana, 51 (5). pp. 1425-1436. ISSN 0126-6039 https://www.ukm.my/jsm/malay_journals/jilid51bil5_2022/KandunganJilid51Bil5_2022.html
institution	Universiti Kebangsaan Malaysia
building	Tun Sri Lanang Library
collection	Institutional Repository
continent	Asia
country	Malaysia
content_provider	Universiti Kebangsaan Malaysia
content_source	UKM Journal Article Repository
url_provider	http://journalarticle.ukm.my/
language	English
description	Cells isolated from skin have wide applications in studies of the pathogenesis of skin-related diseases and the construction of 3D skin equivalents. This study aimed to isolate keratinocytes, fibroblasts, and melanocytes from human foreskin and characterize the purity of the cell types. Keratinocytes, fibroblasts, and melanocytes from human foreskin were isolated by differential trypsinization and media selection. The purity of the cell types was characterized based on the expression of gene markers. The assessment of gene marker expression involved RNA extraction, primer design, quantitative polymerase chain reaction (qPCR) and immunocytochemical staining. Our results showed that in cocultures of keratinocytes and fibroblasts isolated from the dermis, fibroblasts could be separated from keratinocytes by quick trypsinization and culture in Dulbecco’s modified Eagle’s medium. The remaining keratinocytes are cultured in Epilife medium. Melanocytes in cocultures of melanocytes and keratinocytes isolated from the epidermis could be selected by changing Epilife medium to M254 medium. Gene marker results suggested that cytokeratin 14 (CK14) is a suitable marker for keratinocytes, elastin (ELN) is a suitable marker for fibroblasts, and tyrosinase (TYR) and tyrosinase-related proteins 1 and 2 (TYRP1 and TYRP2) are suitable markers for melanocytes. In conclusion, keratinocytes, fibroblasts, and melanocytes can be isolated from the same human foreskin sample by differential trypsinization and media selection techniques and characterized by suitable gene markers. This finding will aid in the isolation of pure skin cell types for various applications in regenerative medicine and toxicity studies.
format	Article
author	Wong, Pau-Ring Nur Fatin Nabilah Mohd Sahardi, Tan, Jen-Kit Chua, Kien-Hui Wan Zurinah Wan Ngah, Suzana Makpol,
spellingShingle	Wong, Pau-Ring Nur Fatin Nabilah Mohd Sahardi, Tan, Jen-Kit Chua, Kien-Hui Wan Zurinah Wan Ngah, Suzana Makpol, Characterization of keratinocytes, fibroblasts and melanocytes isolated from human skin using gene markers
author_facet	Wong, Pau-Ring Nur Fatin Nabilah Mohd Sahardi, Tan, Jen-Kit Chua, Kien-Hui Wan Zurinah Wan Ngah, Suzana Makpol,
author_sort	Wong, Pau-Ring
title	Characterization of keratinocytes, fibroblasts and melanocytes isolated from human skin using gene markers
title_short	Characterization of keratinocytes, fibroblasts and melanocytes isolated from human skin using gene markers
title_full	Characterization of keratinocytes, fibroblasts and melanocytes isolated from human skin using gene markers
title_fullStr	Characterization of keratinocytes, fibroblasts and melanocytes isolated from human skin using gene markers
title_full_unstemmed	Characterization of keratinocytes, fibroblasts and melanocytes isolated from human skin using gene markers
title_sort	characterization of keratinocytes, fibroblasts and melanocytes isolated from human skin using gene markers
publisher	Penerbit Universiti Kebangsaan Malaysia
publishDate	2022
url	http://journalarticle.ukm.my/19477/1/13.pdf http://journalarticle.ukm.my/19477/ https://www.ukm.my/jsm/malay_journals/jilid51bil5_2022/KandunganJilid51Bil5_2022.html
_version_	1743108005678809088
score	13.211869

Characterization of keratinocytes, fibroblasts and melanocytes isolated from human skin using gene markers

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