Human Joint Aging and Inflammatory Studies (I) :Human Articular Chondrocytes Immortalization and Characterization

碩士 === 臺北醫學大學 === 生物醫學材料研究所 === 91 === Articular aging and inflammation has became a tendency of being popular and young. Pathological changes of articular cartilage chondrocytes are the main reason of these diseases. There is, however, still no well-established immortalized articular chondrocyte s...

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Main Authors: Wei-Hong Chen, 陳韋弘
Other Authors: Win-Ping Deng Ph.D.
Format: Others
Language:zh-TW
Published: 2003
Online Access:http://ndltd.ncl.edu.tw/handle/80831922138484950349
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spelling ndltd-TW-091TMC006840022015-10-13T13:35:59Z http://ndltd.ncl.edu.tw/handle/80831922138484950349 Human Joint Aging and Inflammatory Studies (I) :Human Articular Chondrocytes Immortalization and Characterization 人體關節老化與發炎反應研究(I):人體關節軟骨細胞之不朽化與定性 Wei-Hong Chen 陳韋弘 碩士 臺北醫學大學 生物醫學材料研究所 91 Articular aging and inflammation has became a tendency of being popular and young. Pathological changes of articular cartilage chondrocytes are the main reason of these diseases. There is, however, still no well-established immortalized articular chondrocyte system to be used to study the molecular mechanism of articular aging and inflammation. In this experiment, we have successfully established an immortalized human articular chondrocyte transfected with HPV 16 E6/E7 DNA by using retrovirus infection system and this cell line, named hPi, was analyzed and well characterized. RT-PCR, Western blotting, Alcian Blue staining, 3 dimensional cultures and immunohistology were used to demonstrate the characteristic similarity between human articular immortalized and primary chondrocytes. The results showed that the major differentiation marker of articular chondrocytes, type II collagen in mRNA level or protein level in hPi cells were apparently identical with that in primary chondrocytes over 60 passages culture. Lacunae were formed between cells and collagen matrices obviously in hPi cells cultured in type I collagen scaffold in 3 dimension. The GAGs-specific Alcian Blue staining also demonstrated that both primary chondrocytes and hPi cells were stained in blueness. Tumor was not formed on NOD/ SCID mice injected with hPi cells for 3 months. This result indicated that the hPi cell line, chondrocytes immortalized by HPV-16 E6/E7, is non-tumorigenic and could still express cartilage-specific differentiation markers, such as type II collagen and aggrecan through 50 passages in vitro cultures without any redifferentiation strategies. The immortalized human articular chondrocytes will be used to investigate human articular regeneration, cartilage aging and inflammation molecular mechanisms ! Win-Ping Deng Ph.D. 鄧文炳 2003 學位論文 ; thesis 72 zh-TW
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description 碩士 === 臺北醫學大學 === 生物醫學材料研究所 === 91 === Articular aging and inflammation has became a tendency of being popular and young. Pathological changes of articular cartilage chondrocytes are the main reason of these diseases. There is, however, still no well-established immortalized articular chondrocyte system to be used to study the molecular mechanism of articular aging and inflammation. In this experiment, we have successfully established an immortalized human articular chondrocyte transfected with HPV 16 E6/E7 DNA by using retrovirus infection system and this cell line, named hPi, was analyzed and well characterized. RT-PCR, Western blotting, Alcian Blue staining, 3 dimensional cultures and immunohistology were used to demonstrate the characteristic similarity between human articular immortalized and primary chondrocytes. The results showed that the major differentiation marker of articular chondrocytes, type II collagen in mRNA level or protein level in hPi cells were apparently identical with that in primary chondrocytes over 60 passages culture. Lacunae were formed between cells and collagen matrices obviously in hPi cells cultured in type I collagen scaffold in 3 dimension. The GAGs-specific Alcian Blue staining also demonstrated that both primary chondrocytes and hPi cells were stained in blueness. Tumor was not formed on NOD/ SCID mice injected with hPi cells for 3 months. This result indicated that the hPi cell line, chondrocytes immortalized by HPV-16 E6/E7, is non-tumorigenic and could still express cartilage-specific differentiation markers, such as type II collagen and aggrecan through 50 passages in vitro cultures without any redifferentiation strategies. The immortalized human articular chondrocytes will be used to investigate human articular regeneration, cartilage aging and inflammation molecular mechanisms !
author2 Win-Ping Deng Ph.D.
author_facet Win-Ping Deng Ph.D.
Wei-Hong Chen
陳韋弘
author Wei-Hong Chen
陳韋弘
spellingShingle Wei-Hong Chen
陳韋弘
Human Joint Aging and Inflammatory Studies (I) :Human Articular Chondrocytes Immortalization and Characterization
author_sort Wei-Hong Chen
title Human Joint Aging and Inflammatory Studies (I) :Human Articular Chondrocytes Immortalization and Characterization
title_short Human Joint Aging and Inflammatory Studies (I) :Human Articular Chondrocytes Immortalization and Characterization
title_full Human Joint Aging and Inflammatory Studies (I) :Human Articular Chondrocytes Immortalization and Characterization
title_fullStr Human Joint Aging and Inflammatory Studies (I) :Human Articular Chondrocytes Immortalization and Characterization
title_full_unstemmed Human Joint Aging and Inflammatory Studies (I) :Human Articular Chondrocytes Immortalization and Characterization
title_sort human joint aging and inflammatory studies (i) :human articular chondrocytes immortalization and characterization
publishDate 2003
url http://ndltd.ncl.edu.tw/handle/80831922138484950349
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