Comparison of Primary Keloid Fibroblast Cultivation Methods and the Characteristics of Fibroblasts Cultured from Keloids, Keloid-surrounding Tissues, and Normal Skin Tissues

SUMMARY: The establishment of primary keloid fibroblast culture has always been a fundamental measure for studying mechanisms of keloid disease. The quality of the primary cell culture can directly affect the results of further experiments. This study was performed to investigate the optimal growth...

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Autores principales:	Qin,Haiyan, Liu,Ruizhu, Nie,Wenting, Li,Mingxi, Yang,Liehao, Zhou,Changcai, Zhang,Lianbo, Zhang,Guang
Lenguaje:	English
Publicado:	Sociedad Chilena de Anatomía 2021
Materias:	Keloid Normal skin Fibroblasts Primary cell culturing α-SMA
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-95022021000100302
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spelling	oai:scielo:S0717-950220210001003022021-03-01Comparison of Primary Keloid Fibroblast Cultivation Methods and the Characteristics of Fibroblasts Cultured from Keloids, Keloid-surrounding Tissues, and Normal Skin TissuesQin,HaiyanLiu,RuizhuNie,WentingLi,MingxiYang,LiehaoZhou,ChangcaiZhang,LianboZhang,Guang Keloid Normal skin Fibroblasts Primary cell culturing α-SMA SUMMARY: The establishment of primary keloid fibroblast culture has always been a fundamental measure for studying mechanisms of keloid disease. The quality of the primary cell culture can directly affect the results of further experiments. This study was performed to investigate the optimal growth conditions, including the optimal storage time and collagenase treatment time, for in vitro cell culture models and the suitable methods for epidermis-dermis separation in different tissues. Keloid tissues, keloid-surrounding tissues, and normal skin tissues were collected from patients, for primary fibroblast culture. Two methods, tissue explant and collagenase digestion, were deployed and compared. Expression levels of the keloid-related genes α -SMA, Col1, and Col3 were assessed in cells cultured using both methods, to verify the qualities of the primary cells. A comparative analysis was conducted between the two methods and among the three different tissues used. Bacterial and lipid contamination was immediately minimized after the samples were processed. Different methods of epidermis removal and different durations of collagenase digestion were required in different tissues to generate optimal results. Real-time PCR results showed that the mRNA expression levels of keloid-related genes in cultured fibroblasts correlated to their in vivo expression profile, as previously reported in other studies. The results of this study have revealed several key points in the culture of primary keloid fibroblasts and demonstrated the correlation in gene expression between in vivo keloid fibroblasts and in vitro primary keloid fibroblasts.info:eu-repo/semantics/openAccessSociedad Chilena de AnatomíaInternational Journal of Morphology v.39 n.1 20212021-02-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-95022021000100302en10.4067/S0717-95022021000100302
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	Keloid Normal skin Fibroblasts Primary cell culturing α-SMA
spellingShingle	Keloid Normal skin Fibroblasts Primary cell culturing α-SMA Qin,Haiyan Liu,Ruizhu Nie,Wenting Li,Mingxi Yang,Liehao Zhou,Changcai Zhang,Lianbo Zhang,Guang Comparison of Primary Keloid Fibroblast Cultivation Methods and the Characteristics of Fibroblasts Cultured from Keloids, Keloid-surrounding Tissues, and Normal Skin Tissues
description	SUMMARY: The establishment of primary keloid fibroblast culture has always been a fundamental measure for studying mechanisms of keloid disease. The quality of the primary cell culture can directly affect the results of further experiments. This study was performed to investigate the optimal growth conditions, including the optimal storage time and collagenase treatment time, for in vitro cell culture models and the suitable methods for epidermis-dermis separation in different tissues. Keloid tissues, keloid-surrounding tissues, and normal skin tissues were collected from patients, for primary fibroblast culture. Two methods, tissue explant and collagenase digestion, were deployed and compared. Expression levels of the keloid-related genes α -SMA, Col1, and Col3 were assessed in cells cultured using both methods, to verify the qualities of the primary cells. A comparative analysis was conducted between the two methods and among the three different tissues used. Bacterial and lipid contamination was immediately minimized after the samples were processed. Different methods of epidermis removal and different durations of collagenase digestion were required in different tissues to generate optimal results. Real-time PCR results showed that the mRNA expression levels of keloid-related genes in cultured fibroblasts correlated to their in vivo expression profile, as previously reported in other studies. The results of this study have revealed several key points in the culture of primary keloid fibroblasts and demonstrated the correlation in gene expression between in vivo keloid fibroblasts and in vitro primary keloid fibroblasts.
author	Qin,Haiyan Liu,Ruizhu Nie,Wenting Li,Mingxi Yang,Liehao Zhou,Changcai Zhang,Lianbo Zhang,Guang
author_facet	Qin,Haiyan Liu,Ruizhu Nie,Wenting Li,Mingxi Yang,Liehao Zhou,Changcai Zhang,Lianbo Zhang,Guang
author_sort	Qin,Haiyan
title	Comparison of Primary Keloid Fibroblast Cultivation Methods and the Characteristics of Fibroblasts Cultured from Keloids, Keloid-surrounding Tissues, and Normal Skin Tissues
title_short	Comparison of Primary Keloid Fibroblast Cultivation Methods and the Characteristics of Fibroblasts Cultured from Keloids, Keloid-surrounding Tissues, and Normal Skin Tissues
title_full	Comparison of Primary Keloid Fibroblast Cultivation Methods and the Characteristics of Fibroblasts Cultured from Keloids, Keloid-surrounding Tissues, and Normal Skin Tissues
title_fullStr	Comparison of Primary Keloid Fibroblast Cultivation Methods and the Characteristics of Fibroblasts Cultured from Keloids, Keloid-surrounding Tissues, and Normal Skin Tissues
title_full_unstemmed	Comparison of Primary Keloid Fibroblast Cultivation Methods and the Characteristics of Fibroblasts Cultured from Keloids, Keloid-surrounding Tissues, and Normal Skin Tissues
title_sort	comparison of primary keloid fibroblast cultivation methods and the characteristics of fibroblasts cultured from keloids, keloid-surrounding tissues, and normal skin tissues
publisher	Sociedad Chilena de Anatomía
publishDate	2021
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-95022021000100302
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Comparison of Primary Keloid Fibroblast Cultivation Methods and the Characteristics of Fibroblasts Cultured from Keloids, Keloid-surrounding Tissues, and Normal Skin Tissues

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