Hill D J, Crace C J, Fowler L, Holder A T, Milner R D
J Cell Physiol. 1984 Jun;119(3):349-58. doi: 10.1002/jcp.1041190314.
The production of immunologically and biologically active somatomedin activity from isolated myoblasts and fibroblasts from fetal rats of 21 days gestational age was investigated. Myoblast-rich cell populations were derived from primary cultures of dispersed muscle cells by the tendency of myoblasts to become detached from the culture dish in the presence of cytochalasin B. Fibroblasts were obtained from fetal muscle. Culture medium conditioned by exposure to myoblasts for 48 hours produced an increased incorporation of both [35S]sulphate and [3H]thymidine by explants of fetal rat costal cartilage in vitro compared to fresh medium. Myoblast-conditioned medium also contained somatomedin-C-like immunoreactivity which diluted in parallel with partially purified human somatomedin-C (3,271 +/- 446 mU/mg cell protein; mean +/- SEM, seven experiments). Medium conditioned by exposure to fetal rat fibroblasts did not promote isotope uptake by fetal rat cartilage above control values, and contained only low levels of somatomedin-C-like immunoreactivity (343 +/- 89 mU/mg cell protein, three experiments). The release of both somatomedin bioactivity and immunoreactivity into conditioned medium was significantly reduced by the incubation of myoblasts in the presence of rat growth hormone (100 ng/ml and 500 ng/ml). We conclude that fetal rat myoblasts released growth factor activity during culture which exhibited biological and immunologic characteristics of somatomedin. Since the bioactivity was demonstrated on skeletal tissues from rat fetuses of the same gestational age as those that yielded myoblasts such growth factor release may be physiological.
对来自妊娠21天的胎鼠的分离成肌细胞和成纤维细胞产生免疫和生物活性生长调节素活性进行了研究。富含成肌细胞的细胞群体是通过在细胞松弛素B存在下成肌细胞从培养皿上脱离的趋势,从分散的肌肉细胞原代培养物中获得的。成纤维细胞取自胎儿肌肉。与新鲜培养基相比,经成肌细胞处理48小时的培养基能使胎鼠肋软骨外植体在体外对[35S]硫酸盐和[3H]胸腺嘧啶核苷的摄取增加。成肌细胞条件培养基还含有生长调节素C样免疫反应性,其稀释情况与部分纯化的人生长调节素C平行(3,271±446 mU/mg细胞蛋白;平均值±标准误,七次实验)。经胎鼠成纤维细胞处理的培养基不会使胎鼠软骨对同位素的摄取高于对照值,且仅含有低水平的生长调节素C样免疫反应性(343±89 mU/mg细胞蛋白,三次实验)。在大鼠生长激素(100 ng/ml和500 ng/ml)存在下培养成肌细胞,可显著降低生长调节素生物活性和免疫反应性向条件培养基中的释放。我们得出结论,胎鼠成肌细胞在培养过程中释放生长因子活性,该活性表现出生长调节素的生物学和免疫学特征。由于这种生物活性在与产生成肌细胞的胎鼠相同胎龄的骨骼组织上得到证实,因此这种生长因子的释放可能是生理性的。