Isolating human dermal fibroblasts using serial explant culture.

Q1 Biochemistry, Genetics and Molecular Biology
Fereshte Nejaddehbashi, V. Bayati, L. Mashali, M. Hashemitabar, M. Abbaspour, E. Moghimipour, M. Orazizadeh
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引用次数: 11

Abstract

Background The purpose of this study was to introduce an applicable culture technique to isolate human dermal fibroblasts (HDFs); which could also contribute to research, clinical practices, as well as tissue engineering. Methods Samples from the human skin were dissected and cultured via serial explant technique. Subsequently, the isolated fibroblasts were assessed for their protein markers and genetic variations via immunofluorescence (IF) and karyotyping; respectively. Following the employment of this technique, a small piece of explant completely disappeared; while no dermis remained after 10 days. Results The quantity of HDFs harvested through this culture technique was reported at a normal level. The results of immunostaining also indicated that the isolated fibroblasts had expressed vimentin and fibronectin; whereas no cells had shown cytokeratin and epidermal marker. Moreover, karyotyping results for the fibroblasts isolated by the given technique revealed no chromosomal diversity after passage 20. Conclusions It was concluded that serial explant culture was an efficient technique for isolating HDFs from a small piece of skin in short-time periods; which could also preserve their normal morphology and molecular characteristics.
使用连续外植体培养分离人真皮成纤维细胞。
本研究的目的是介绍一种分离人真皮成纤维细胞(HDFs)的适用培养技术;这也有助于研究,临床实践,以及组织工程。方法采用连续外植体技术对人体皮肤进行解剖培养。随后,分离的成纤维细胞通过免疫荧光(IF)和核型评估其蛋白质标记和遗传变异;分别。采用这种技术后,一小块外植体完全消失;10天后,真皮不复存在。结果通过该培养技术获得的HDFs数量处于正常水平。免疫染色结果还表明,分离的成纤维细胞表达了波形蛋白和纤维连接蛋白;而未见细胞角蛋白和表皮标记。此外,用该技术分离的成纤维细胞的核型结果显示,在传代20次后,没有染色体多样性。结论连续外植体培养可在短时间内从小块皮肤中分离出HDFs;这也可以保持它们的正常形态和分子特征。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Stem cell investigation
Stem cell investigation Biochemistry, Genetics and Molecular Biology-Developmental Biology
CiteScore
5.80
自引率
0.00%
发文量
9
期刊介绍: The Stem Cell Investigation (SCI; Stem Cell Investig; Online ISSN: 2313-0792) is a free access, peer-reviewed online journal covering basic, translational, and clinical research on all aspects of stem cells. It publishes original research articles and reviews on embryonic stem cells, induced pluripotent stem cells, adult tissue-specific stem/progenitor cells, cancer stem like cells, stem cell niche, stem cell technology, stem cell based drug discovery, and regenerative medicine. Stem Cell Investigation is indexed in PubMed/PMC since April, 2016.
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