Shinji T, Koide N, Tsuji T
First Department of Internal Medicine, Okayama University Medical School, Japan.
Cell Struct Funct. 1988 Apr;13(2):179-88. doi: 10.1247/csf.13.179.
Adult rat hepatocytes seeded in a noncoated plastic dish containing serum-free medium formed a monolayer within 24 h of culture. Those seeded in a dish coated with a proteoglycan fraction isolated from rat liver reticulin fibers attached to the dish but did not spread within 4 h, and then gradually assembled to form floating spherical aggregates (spheroids) with a diameter of 120 +/- 40 micron, within 72 h. The proteoglycan fraction appeared to contain dermatan sulfate, heparan sulfate and an unidentified glycosaminoglycan in its glycan moieties by glycosaminoglycan analysis after pronase digestion and high molecular weight proteoglycan molecules (mw: over 300,000 and about 200,000) by SDS-PAGE analysis. Cells seeded in dishes coated with these defined glycosaminoglycans and heparin assembled to form hemispheroids and multilayer islands, but not floating spheroids, within 72 h of culture. Dermatan sulfate had a stronger ability to induce hemispheroids than heparan sulfate or heparin. As the hemispheroid and multilayer islands were the intermediate form between monolayer and floating spheroids, the glycosaminoglycan moieties of the proteoglycan fraction were thought to participate in the formation of spheroid.
接种于含无血清培养基的未包被塑料培养皿中的成年大鼠肝细胞，在培养24小时内形成单层。接种于涂有从大鼠肝网硬蛋白纤维中分离出的蛋白聚糖组分的培养皿中的肝细胞，在4小时内附着于培养皿但未铺展，然后在72小时内逐渐聚集形成直径为120±40微米的漂浮球形聚集体（球体）。经链霉蛋白酶消化后的糖胺聚糖分析表明，该蛋白聚糖组分的聚糖部分似乎含有硫酸皮肤素、硫酸乙酰肝素和一种未鉴定的糖胺聚糖，通过SDS-PAGE分析显示含有高分子量的蛋白聚糖分子（分子量：超过300,000和约200,000）。接种于涂有这些确定的糖胺聚糖和肝素的培养皿中的细胞，在培养72小时内聚集形成半球体和多层岛状物，但未形成漂浮球体。硫酸皮肤素诱导半球体形成的能力比硫酸乙酰肝素或肝素更强。由于半球体和多层岛状物是单层和漂浮球体之间的中间形式，因此认为该蛋白聚糖组分的糖胺聚糖部分参与了球体的形成。
