Tschan T, Höerler I, Houze Y, Winterhalter K H, Richter C, Bruckner P
Laboratorium für Biochemie, Eidgenössische Technische Hochschule, Zürich, Switzerland.
J Cell Biol. 1990 Jul;111(1):257-60. doi: 10.1083/jcb.111.1.257.
Chondrocytes in dense suspension culture in agarose survive in serum-free DME because they secrete low molecular mass compounds supporting their own viability. This activity can be replaced by pyruvate, or sulfhydryl compounds, e.g., cysteine or dithioerythritol. Catalase, an enzyme decomposing H2O2, also protects the cells, whereas superoxide dismutase has no effect. Therefore, chondrocytes in culture are sensitive to toxic compounds derived from molecular oxygen, i.e., hydroxyl radicals or hydrogen peroxide spontaneously generated in DME containing ascorbate and ferrous ions. Poly-ADP-ribosylation is an important step in the cascade of events triggered by these compounds. To survive, chondrocytes do not require stimulation by growth factors. They remain resting cells in fully defined, serum-free culture also at low density. Proliferation and hypertrophy can be induced by serum, but not by low cell density alone.
琼脂糖中密集悬浮培养的软骨细胞能在无血清的杜氏改良伊格尔培养基(DME)中存活,因为它们分泌支持自身生存能力的低分子量化合物。这种活性可以被丙酮酸或巯基化合物(如半胱氨酸或二硫苏糖醇)所替代。过氧化氢酶是一种分解过氧化氢的酶,它也能保护细胞,而超氧化物歧化酶则没有作用。因此,培养中的软骨细胞对分子氧衍生的有毒化合物敏感,即在含有抗坏血酸盐和亚铁离子的DME中自发产生的羟基自由基或过氧化氢。多聚ADP-核糖基化是由这些化合物触发的一系列事件中的重要一步。为了存活,软骨细胞不需要生长因子的刺激。在完全确定的无血清培养中,即使细胞密度很低,它们也保持静止状态。血清可诱导细胞增殖和肥大,但仅低细胞密度不能诱导。