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    • 5. 发明申请
    • METHOD OF PRODUCTION OF TRANSGENIC AVIAN USING EMBRYONIC STEM CELLS
    • 使用胚胎干细胞生产转基因禽的方法
    • WO2008017704A1
    • 2008-02-14
    • PCT/EP2007/058263
    • 2007-08-09
    • VIVALISVALARCHE, IsabelleBATARD, LucMEHTALI, Majid
    • VALARCHE, IsabelleBATARD, LucMEHTALI, Majid
    • C12N5/06
    • C12N5/0606A01K2217/00A01K2227/30C12N2501/10C12N2501/105C12N2501/115C12N2501/125C12N2501/13C12N2501/23C12N2501/2306C12N2502/13
    • The present invention concerns a method of culturing embryonic stem (ES) cells of avian, comprising the steps of: a) suspending ES cells originating from the blastoderm disk of fertilized un-incubated avian egg(s) in a basal culture medium supplemented with: insulin-like growth factor-1 (IGF-1) and ciliary neurotrophic factor (CNTF); and animal serum; and optionally, at least one growth factors selected in the group comprising interleukin 6 (Il-6), interleukin 6 receptor (Il-6R), stem cell factor (SCF), fibroblast growth factor (FGF), leukaemia inhibitory factor (LIF), interleukin 11 (Il-11), oncostatin and cardiotrophin; b) seeding the suspension of ES cells obtained in step a) on a layer of feeder cells and further culturing the ES cells for at least between 2 to 10 passages; c) optionally removing at least one growth factors selected SCF, FGF, Il-6, Il-6R, LIF, oncostatin and cardiotrophin and Il-11 from the culture medium; d) further culturing the ES cells in the medium of step c) on a layer of feeder cells.
    • 本发明涉及一种培养禽类胚胎干(ES)细胞的方法,包括以下步骤:a)将源自受精培养未孵化禽鸟卵胚细胞盘的ES细胞悬浮在补充以下物质的基础培养基中: 胰岛素样生长因子-1(IGF-1)和睫状神经营养因子(CNTF); 和动物血清; 并且任选地,选自包含白细胞介素6(II-6),白细胞介素6受体(II-6R),干细胞因子(SCF),成纤维细胞生长因子(FGF),白血病抑制因子(LIF))的组中的至少一种生长因子, ,白介素11(Il-11),制瘤素和心肌营养素; b)将步骤a)中获得的ES细胞的悬浮液接种在饲养细胞层上,并进一步培养ES细胞至少2至10代; c)任选地从培养基中除去选自SCF,FGF,II-6,II-6R,LIF,制瘤素和心肌营养素和II-11的至少一种生长因子; d)在步骤c)的培养基中进一步培养ES细胞在饲养细胞层上。