- 期刊
以中國倉鼠卵巢細胞表現具生物活性之豬顆粒球巨噬細胞群落刺激因子重組蛋白
Expression of Biologically Active Porcine Granulocytel Macrophage Colony-Stimulating Factors (GM-CSF) by Chinese Hamster Ovary (CHO) Cells
摘要
群落刺激因子(colony-stimulating factors; CSFs)家族包含巨噬細胞群落刺激因子(macrophage-CSF; MCSF)、顆粒球群落刺激因子(granulocyte-CSF; G-CSF)、顆粒球巨噬細胞群落刺激因子(granulocyte/macrophage-CSF; GM-CSF)及多群落刺激因子(multi-CSF; interleukin 3)等，是一群調控血球細胞造血作用的重要細胞激素。GM-CSF目前已廣泛應用於試管內誘發骨髓造血母細胞與週邊血液單核球增生分化為樹狀突細胞之細胞激素。本實驗產生具有穩定表現豬GM-CSF之中國倉鼠卵巢細胞株，所表現之豬GM-CSF定序確認與GenBank AY116504序列完全相同，以硫酸十二酯鈉－聚丙烯醯胺膠體電泳(sodium dodecyl sulfate-polyacrylamide gel electrophoresis; SDSPAGE)及西方轉漬法確認細胞培養液中含豬GM-CSF重組蛋白，分子量為17.6kDa，以豬骨髓造血母細胞群落生物活性檢測，細胞培養液中豬GM-CSF重組蛋白之生物活性為3000U/mL。
並列摘要
The colony-stimulating factors (CSFs) consisting of the macrophage-CSF (M-CSF), granulocyte-CSF (G-CSF), granulocyte/macrophage-CSF (GM-CSF), and multi-CSF (interleukin 3) are essential to the hematopoiesis of blood cells. GM-CSF is one of the most important stimulatory cytokines for in vitro proliferation and differentiation of bone marrow hematopoietic cells or peripheral blood mononuclear cells into functional dendritic cells. In the present study, porcine GM-CSF (pGM-CSF) was expressed using Chinese hamster ovary cells (CHO) as the expression system. The molecular weight of the expressed recombinant pGM-CSF was 17.6kD identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. The biological activity of pGM-CSF in culturing medium of the stable clone of CHO cells was 3000U/mL using the colony bioassay of swine bone marrow haematopoietic cells.