| Summary: | 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 !
|
|---|
