胶质瘤动物模型课件

,单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,胶质瘤动物模型,Glioma Animal model,1、实验诱发模型,1.1 化学诱发,1.2 病毒诱发,2、移植瘤模型,2.1 原位移植瘤,2.2 皮下移植瘤,3、转基因小鼠模型,Glioma Animal model,首见于 1939 年 Seligman 把多环芳香烃类的甲基胆蒽制成丸剂植入小鼠脑内，诱发成胶质瘤和肉瘤。,从上世纪中叶起，因利用亲神经的烷化剂全身给药诱发神经系统肿瘤获得成功。,甲基亚硝基脲和乙基亚硝基脲有很高的中枢神经系统肿瘤诱发率，至今还在运用的 P494、C6、9L 和G422 等胶质瘤动物模型都是用上述类似方法制作成功,。,1、实验诱发模型化学诱发,病毒诱发有两类，一是核糖核酸病毒，如劳斯肉 瘤 病 毒；二 是 脱 氧 核 糖 核 酸 病 毒，如 腺 病 毒。,Bigner 等把浓缩的呼吸道合胞病毒（Respiratory Syncytial Virus，RSV）注射到新生犬脑内，经过一段潜伏期后全部发生脑胶质瘤。,1、实验诱发模型病毒诱发,已建系的肿瘤细胞株,腹水瘤移植接种法,实体瘤移植接种法（,1,）小块接种法（,2,）肿瘤组织浆接种法,2、移植瘤模型皮下移植瘤,移植瘤的接种方法,2、移植瘤模型皮下移植瘤,（李承科，霍刚，郑履平，等.快速建立胶质瘤模型的研究，重庆医科大学学报J.2008,33(1):73-76）,C6/SD rat,C6/Wistar rat,9L/F344 rat,CNS-1/Lewis rat,U87MG/,Swiss-nu,U87/BALB/nu-c,2、移植瘤模型原位移植瘤,靶点坐标：大鼠右侧尾状核,区，前囟中点前1.0mm，矢状缝右旁开3.0mm，硬脑膜下5mm,2、移植瘤模型原位移植瘤,Implantation technique,Anesthesia was induced by inhalation of a mixture of oxygen with 5%halothane,followed by an intra-peritoneal injection of ketamine(87 mg/kg)and xylazine(13 mg/kg)for maintenance.Animals were then mounted on a stereotactic frame.Special care was used in the placement of the head,in the frame to avoid sagittal angulation,so that the head was,parrallel to the frame stand.A midline scalp incision was performed,followed by identification and exposure of the bregma.Using a 16-gauge needle,a burr hole was placed on the right frontal bone.A 25-,microliter SGF syringe with a 27-gauge needle secured to the frame was used to infuse the cellular solution over a period of five minutes.The needle was then slowly withdrawn.Bone wax was applied to close the burr hole and the scalp was closed with a continuous one-layer resorbable suture.,David Mathieu,Roger Lecomte,Ana Maria Tsanaclis,et al.Standardization anddetailed characterization of the syngeneic Fischer/F98 glioma model J.,LE JOURNAL CANADIEN DES SCIENCES NEUROLOGIQUES,2007,34(3):296-306),C6细胞,是经N一亚硝基甲脲诱发的大鼠脑胶质瘤细胞，来源于远交系Wistar大鼠。C6细胞MHC中存在与BDIX、BDX、SD、Wistar等种系大鼠异源性基因，C6/Wistar模型中，肿瘤及瘤周淋巴细胞浸润和体液抗胶质瘤抗体反应均明显高于9L/F344，显示C6细胞自身有很强免疫原性。模型中因MHC差异引发的免疫排斥反应，可能造成脑内肿瘤不能持续增长。非免疫缺陷C6荷瘤大鼠模型用于脑胶质瘤免疫或基因免疫治疗研究，其自身存在的免疫排斥反应可能混淆治疗所产生效果。故建议采用同源基因型的9L/F344大鼠脑胶质瘤模。,9L细胞,是由近交系F344大鼠经N一亚硝基甲脲体内诱发的恶性脑胶质肉瘤细胞，其主要组织相容性基因复合体(MHC)与F344大鼠同源性高。,（林健,王伟民,徐如祥,等.9L/F344与C6/Wistar大鼠脑胶质瘤模型局部细胞免疫反应的比较研究J.2004,9(9):409-412）,Purpose:High-grade gliomas are closely related to the mesenchymal phenotype which might be explained by unorthodox differentiation of glioma cancer stem cells,(gCSCs).We reasoned that other non-neural stem cells,especially mesenchymal stem cells(MSCs),might play a role in expresssing mesenchymal phenotype of high-grade,gliomas.Thus we hypothesized that cells resembling MSCs,exist in glioma specimens.,Presence of glioma stroma mesenchymal stem,Cells in a murine orthotopic glioma model,Sang-Mok Kim&Seok-Gu Kang&Na-Ri Park,et al.Springer-Verlag 2011,Methods:We created a mouse(m)orthotopic glioma model,using human gCSCs.Single-cell suspensions were isolated,from glioma specimens and cultured according to the,methods for mMSCs or gliomaspheres.These cells were,analyzed by fluorescence-activated cell sorting(FACS)for,surface markers associated with mMSCs or gCSCs.Glioma,stroma(GS)-MSCs were exposed to mesenchymal differ-,entiation conditions.To decide the location of GS-MSCs,sections of orthotopic glioma models were analyzed by,immunofluorescent labeling.,Presence of glioma stroma mesenchymal stem,Cells in a murine orthotopic glioma model,Presence of glioma stroma mesenchymal stem,Cells in a murine orthotopic glioma model,Presence of glioma stroma mesenchymal stem,Cells in a murine orthotopic glioma model,Presence of glioma stroma mesenchymal stem,Cells in a murine orthotopic glioma model,Presence of glioma stroma mesenchymal stem,Cells in a murine orthotopic glioma model,遗传学控制分类,实验动物的分类,1、近交系,(,inbred strain,),2、封闭群(远交群,)(closed,colony),3、杂交群(hybrids),4、突变系(mutant strain),5、转基因动物(transgenic animals),1、近交系,(inbred strain),近交系,经至少连续20代全同胞兄妹交配（或者亲代与子代交配）培育而成，品系内所有个体都可追溯到起源于第20代或以后代数的一对共同祖先。,命名,一般以大写英文字母或大写英文字母加阿拉伯数字命名，如,DBA、C3H,等，一般在品系符号后的括号里用F加上数字指示其兄妹交配的代数，如,C3H（F130）,。,DBA,封闭群,以非近亲交配方式进行繁殖生产，在不引入外部新个体的条件下，至少连续繁殖4代以上，称为一个封闭群，或叫远交群。如：,NIH,小鼠、,Wistar,大鼠和新西兰兔等。,2、封闭群(closed colony),命名,保持者英文缩写名称:种群名称组成。如,N:NIH,表示由美国卫生研究院(National institutes of Health,N)保持的NIH封闭群小鼠。,Lac:LACA,表示由英国实验动物中心(Lac)保持LACA封闭群小鼠。,某些命名较早且又广为人知的封闭群动物，仍沿用其原来的名称。如,Wistar,大鼠。,2、封闭群(closed colony),NIH,Wistar,3、杂交群（hybrids）,杂交群,不同品系或种群之间杂交产生的后代。,命名,雌性亲代名称X雄性亲代名称，在后面加上杂交代数。,如：,C57BL/6XDBA/2F1,表示以,C57BL/6,为母系，,DBA/2,为父系杂交生育的杂交一代，可简写为,B6D2F1,。,突变系,受各种内外因素影响引起染色体畸变和基因突变而育成某些特殊性状表型的品系。,命名,在其它群命名基础上加上突变基因符号，用连字号相连，基因符号用斜体字。如：,NIH-,nu,/,nu,表示由美国国立卫生研究院保存的,带有纯合裸基因的NIH小鼠。,4、,突变系,(mutant strain),Nature:,雌性也疯狂,转基因动物,通过实验手段把外源基因稳定地整合至基因组，并能遗传给后代的动物。,命名,C57BL/6J-TgN(CD8Ge)23Jwg,5、,转基因动物,(,transgenic animals),Thank you for your attention!,