人表皮细胞培养适宜条件的探讨

发布时间：2018-04-30 10:57

本文选题：表皮细胞分离液 + 筛网过滤　；参考：《郑州大学》2006年硕士论文

【摘要】：背景表皮细胞培养用于探讨表皮生物学特性以及多种皮肤病的发病机理，也可用于皮肤药理学，在烧伤外科及整形外科中亦有着广阔的前景。人类进行表皮细胞体外培养已有100余年的历史，，很长时间内不能解决人表皮细胞的体外长期大量培养问题，直到1975年Rheinwald和Green利用放射致死的3T3鼠成纤维细胞作为表皮细胞培养的滋养层细胞，才使表皮细胞的体外长期培养扩增成为可能。随着现代分子生物学和组织工程学的迅猛发展，以及研究手段和实验条件的进一步改善，表皮角质形成细胞培养在理论和技术上都有了不断的发展和完善。关于表皮细胞体外培养的适宜方法和条件，国内外已有不少报道。然而，关于表皮细胞的分离液成分方面对表皮的克隆形成率，融合天数等细胞生物学性质的影响却缺乏深入的对比研究。为建立适宜的稳定培养条件，包括滋养层问题，培养液内容问题，尤其是关于表皮细胞分离液成分等问题，我们对正常人表皮生物工程中的适宜条件进行了探讨。此外，为了获得更多的表皮深层细胞，我们还对分离表皮后的表皮细胞通过不同孔径的尼龙布纱网过滤，对比不同的尼龙布纱网滤过的表皮细胞中深层细胞(主要指表皮干细胞)与中、浅层细胞的比例，为有关表皮生物学进一步提供理论资料并为表皮生物工程研究奠定实验基础。材料与方法标本均取自我院2005年5月-2005年8月泌尿外科门诊及住院包皮过长患者4例。年龄24-35岁。经各项检查证实无心、肝、肺、内分泌、神经系统疾病及肿瘤。以NIH-3T3细胞作为滋养层，以热溶素分离表皮与真皮，分离表皮细胞应用
[Abstract]:Background The culture of epidermal cells can be used to study the biological characteristics of epidermis and the pathogenesis of various skin diseases. It can also be used in dermatopharmacology and has a broad prospect in burn surgery and plastic surgery. Human epidermal cells have been cultured in vitro for more than 100 years, which can not solve the problem of human epidermal cells culture in vitro for a long time. It was not until 1975 that Rheinwald and Green used lethal 3T3 fibroblasts as trophoblast cells cultured in epidermal cells, which made it possible to expand epidermal cells in vitro for a long time. With the rapid development of modern molecular biology and tissue engineering, as well as the further improvement of research methods and experimental conditions, epidermal keratinocyte culture has been continuously developed and improved in theory and technology. There have been many reports on the suitable methods and conditions of epidermal cell culture in vitro. However, there is a lack of in-depth comparative study on the effects of the components of the separation fluid of epidermal cells on the cell biological properties such as the formation rate of epidermis, the days of fusion and so on. In order to establish suitable stable culture conditions, including trophoblast problem, culture medium content, especially the composition of epidermal cell isolate, we discussed the suitable conditions in normal human epidermis bioengineering. In addition, in order to obtain more deep epidermal cells, we also filter the epidermal cells after the separation of the epidermis through nylon gauze mesh with different pore sizes. The ratio of the deep cells (mainly epidermal stem cells) to the superficial cells (mainly epidermal stem cells) in the epidermal cells filtered by different nylon gauze mesh was compared, which provided further theoretical data for epidermal biology and laid an experimental foundation for the study of epidermal bioengineering. Materials and methods The specimens were collected from 4 outpatients of urology outpatient and inpatients in our hospital from May 2005 to August 2005. They are 24-35 years old. Unintentional, liver, lung, endocrine, nervous system diseases and tumors were confirmed by various examinations. NIH-3T3 cells were used as trophoblast, epidermis and dermis were separated by thermolysin, and epidermal cells were separated.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R329.2

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