摘要
Previously, we described the genetic construction Super- CHO, a cell line capable of autocrine growth under fully defined protein-free conditions. Super-CHO cells constitutively express insulin growth factor-I (IGF-I) and transferrin in sufficient amounts to support long-term, stable growth without the addition of exogenous growth factors, thus making it an ideal host for the production of recombinant biopharmaceuticals. although IGF-I has been successfully expressed in Chinese Hamster Ovary (CHO) cells, the long term effects of recombinant IGF-I expression have not been explored. In particular, the expression of the endogenous IGF-I receptor in response to IGF-I production has not been reported. We report here the transcriptional induction of the type I IGF receptor gene in Super-CHO. In addition, we examined the conditioned medium for the presence of IGF-I binding proteins. Ligand blot analysis reveals the presence of IGF binding proteins present in the medium conditioned by Super-CHO cells as well as CHO cells incubated in the presence of IGF-I. Furthermore, immunoaffinity reveals that Super-CHO expresses IGF binding protein-3 in response to IGF-I production. These results suggest the autocrine growth of Super-CHO involves a complex interaction of cell type specific factors which regulate its utility of IGF-I. autocrine growthCHOIGF-IIGFBPIGF-I receptor
摘要译文
以前，我们描述了基因构建Super-CHO，一种能够在完全确定的无蛋白条件下自分泌生长的细胞系超级CHO细胞组成型表达足够量的胰岛素生长因子-I（IGF-I）和转铁蛋白，以支持长期稳定生长而不添加外源性生长因子，从而使其成为生产重组生物药物的理想宿主，尽管IGF-1已经在中国仓鼠卵巢（CHO）细胞中成功表达，但尚未探索重组IGF-1表达的长期效应。特别地，尚未报道响应IGF-1产生的内源性IGF-1受体的表达我们在这里报告超级CHO中的I型IGF受体基因的转录诱导此外，我们检测条件培养基中IGF-I结合蛋白的存在配体印迹分析揭示存在于由Super-CHO细胞以及在IGF-I the存在下孵育的CHO细胞条件化的培养基中的IGF结合蛋白的存在。此外，免疫亲和显示超级CHO表达IGF结合蛋白-3响应IGF-1生产这些结果表明Super-CHO的自分泌生长涉及细胞类型特异性因子的复杂相互作用，其调节其对IGF-1自分泌生长CHOIGF-IIGFBPIGF-1受体的效用
Noelle-Anne Sunstrom [1] ;Masood Baig [2] ;Louise Cheng [3] ;Derick Payet Sugyiono [3] ;Peter Gray [3]. Recombinant insulin-like growth factor-I (IGF-I) production in Super-CHO results in the expression of IGF-I receptor and IGF binding protein 3[J]. Cytotechnology, 1998,28(1-3): 91–100