Centrella M, Canalis E
Proc Natl Acad Sci U S A. 1985 Nov;82(21):7335-9. doi: 10.1073/pnas.82.21.7335.
Conditioned medium recovered from fetal rat calvarial cultures contains an autocrine factor termed bone-derived growth factor (BDGF); this factor has been purified by acid extraction, gel-permeation chromatography, and two reversed-phase HPLC steps and examined for mitogenicity on normal rat kidney fibroblasts (NRK, clone 49F). HPLC-purified BDGF caused a dose-related increase in cell number, DNA content, and [3H]thymidine incorporation into acid-insoluble material. Since highly purified BDGF appeared less mitogenic than cruder preparations, the latter were tested for additional growth factors, with particular attention to those required for anchorage-independent colony formation in soft agar. BDGF did not displace 125I-labeled epidermal growth factor (EGF) in a radioligand-receptor assay, indicating the absence of EGF and transforming growth factor alpha (TGF-alpha). Without EGF, no BDGF preparation induced NRK cells to form soft agar colonies. However, calvarial conditioned medium contained a factor which, like TGF-beta, induced large soft-agar colonies in the presence of EGF; this TGF-beta-like factor did not copurify with BDGF. Polyclonal antibodies against platelet-derived growth factor did not neutralize the effects of BDGF on NRK cells. BDGF is a potent mitogen for nonskeletal-tissue-derived fibroblasts. Although crude BDGF preparations do contain TGF-beta, BDGF is distinct from this factor and others necessary for NRK cell transformation to anchorage-independent growth.
从胎鼠颅骨培养物中回收的条件培养基含有一种自分泌因子，称为骨衍生生长因子（BDGF）；该因子已通过酸提取、凝胶渗透色谱和两步反相高效液相色谱法进行纯化，并检测了其对正常大鼠肾成纤维细胞（NRK，克隆49F）的促有丝分裂活性。高效液相色谱法纯化的BDGF导致细胞数量、DNA含量以及[3H]胸苷掺入酸不溶性物质的量呈剂量相关增加。由于高度纯化的BDGF似乎比粗制品的促有丝分裂活性低，因此对后者进行了其他生长因子的检测，尤其关注软琼脂中不依赖贴壁的集落形成所需的因子。在放射性配体-受体分析中，BDGF不能取代125I标记的表皮生长因子（EGF），表明不存在EGF和转化生长因子α（TGF-α）。没有EGF时，没有BDGF制剂能诱导NRK细胞形成软琼脂集落。然而，颅骨条件培养基含有一种因子，与TGF-β一样，在EGF存在下能诱导形成大的软琼脂集落；这种TGF-β样因子与BDGF不能共同纯化。抗血小板衍生生长因子的多克隆抗体不能中和BDGF对NRK细胞的作用。BDGF是一种对非骨骼组织来源的成纤维细胞有效的促有丝分裂原。虽然粗制的BDGF制剂确实含有TGF-β，但BDGF与该因子以及NRK细胞转化为不依赖贴壁生长所需的其他因子不同。
