@article{e0793d8626fa4e9789e79fa866a9f8ba,
title = "Coordinated change between complement C1s production and chondrocyte differentiation in vitro",
abstract = "In vitro synthesis of the first component of complement C1s was examined by using hamster epiphyseal chondrocytes (HAC) and human chondrosarcoma cell line HCS-2/8. Hamster and human C1s produced by the cells were quantified by immunoblotting and sandwich enzyme-linked immunosorbent assay (ELISA), respectively. It was possible to measure active and inactive C1s by sandwich ELISA, when we used anti-human C1s monoclonal antibodies, M241 recognizing only active C1s, and M365 and M81 recognizing both active and inactive C1s. Approximately 40% of C1s secreted from HCS-2/8 was found to be activated in the culture medium, whereas C1s from HAC was not. C1s production increased in accordance with chondrocyte differentiation induced by ascorbic acid. In contrast, transforming growth factor-beta1 and basic fibroblast growth factor, which inhibited differentiation, suppressed C1s production. These results confirmed our previous observation showing that C1s synthesis increased with differentiation into hypertrophic chondrocytes in vivo.",
keywords = "Ascorbic acid, Cell culture, Chondrocytes, Complement C1s, Differentiation, Human, Sandwich ELISA, Syrian hamster",
author = "Koichi Nakagawa and Hisako Sakiyama and Takeshi Fukazawa and Misako Matsumoto and Masaharu Takigawa and Toru Toyoguchi and Hideshige Moriya",
note = "Funding Information: In order to obtain hamster chondrocytes (HAC), epiphyseal cartilage of knees and shoulders of 3-day-old Syrian hamsters was digested with 0.2% collagenase (type IV, Sigma Chemical, St. Louis, Mo., USA) in minimal essential medium (MEM) plus 10% fetal calf serum (FCS) for 3 h. HAC were cultured in a 1:1 mixture of Dulbecco{\textquoteright}s modified Eagle{\textquoteright}s medium and F-12 (DMEM+F-12), supplemented with 20% FCS. Secondary cultures of HAC were used for experiments throughout. Cells of human chondrosarcoma cell line HCS-2/8 were maintained in MEM supplemented with 20% FCS. At various cell densities, viz., sparse, confluent, and over-confluent, the medium was switched to serum-free collection medium, DMEM+F-12, or MEM supplemented with human transferrin (10 µg/ml), bovine insulin (10 µg/ml), and hydrocortisone (10–8 M). After a 24-h incubation, the collection medium was harvested and assayed for type II collagen and C1s content. Nil2C2 cells, a malignant cell line derived from ham- This work was supported in part by Special Coordination Funds for Promoting Science and Technology from the Science and Technology Agency of Japan. Correspondence to: H. Sakiyama (Tel: +81–43–251–2111 ext. 456; Fax: +81–43–251–4582)& ster embryo fibroblasts (Sakiyama and Robbins 1973), were maintained in DMEM+F-12, supplemented with the three growth factors described above.",
year = "1997",
doi = "10.1007/s004410050876",
language = "English",
volume = "289",
pages = "299--305",
journal = "Zeitschrift für Zellforschung und mikroskopische Anatomie (Vienna, Austria : 1948)",
issn = "0302-766X",
publisher = "Springer Verlag",
number = "2",
}
