Transforming and nontransforming growth factors are present in medium conditioned by fetal rat calvariae.
Transforming and nontransforming growth factors are present in medium conditioned by fetal rat calvariae.
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转化和非转化生长因子存在于由胎鼠颅盖调节的培养基中。
DOI：
10.1073/pnas.82.21.7335
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发表时间：
1985
影响因子：
11.1
通讯作者：
Canalis,E
中科院分区：
文献类型：
--
作者：
Centrella,M;Canalis,E
从胎鼠颅骨培养物中回收的条件培养基含有一种称为骨源性生长因子（BDGF）的自分泌因子;该因子已通过酸提取、凝胶渗透色谱法和两个反相HPLC步骤进行纯化，并检查了对正常大鼠肾成纤维细胞（NRK，克隆49 F）的促有丝分裂作用。HPLC纯化的BDGF引起细胞数量、DNA含量和[3 H]胸苷掺入酸不溶性物质的剂量相关性增加。由于高度纯化的BDGF比粗品制剂的促有丝分裂性更低，因此对后者进行了额外生长因子的测试，特别注意软琼脂中锚定非依赖性菌落形成所需的生长因子。在放射性配体-受体测定中，BDGF没有取代125 I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，没有BDGF制剂诱导NRK细胞形成软琼脂集落。然而，颅骨条件培养基中含有一个因子，如TGF-β，诱导大的软琼脂菌落在EGF的存在下，这种TGF-β样因子不与BDGF共纯化。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是非乳腺组织来源的成纤维细胞的有效有丝分裂原。尽管BDGF粗品确实含有TGF-β，但BDGF与该因子以及NRK细胞转化为非贴壁依赖性生长所必需的其他因子不同。
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.