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hCSF2 NPG 小鼠

1. 基本信息

品系名称 NOD. Prkdcscid Il2rgtm1 Vst Tg(Csf1p – hCSF2) / Vst

常用名  hCSF2-NPG /Vst; hCSF2 NPG

背景    NOD-scid/NcrCrl

毛色    白色


品系建立

采(cai)用转基因的(de)方法,扩增(zeng)小(xiao)鼠CSF1的(de)ATG上(shang)游(you)1.2Kb的(de)片(pian)(pian)段(duan)(含(han)启动子),与人CSF2的(de)CDS连接,插入(ru)pCDNA3.1载(zai)(zai)体中(zhong),构建(jian)pCDNA3.1-Csf1p-hCSF2表(biao)达载(zai)(zai)体(如下(xia)图所示)。酶切(qie)分离表(biao)达载(zai)(zai)体片(pian)(pian)段(duan),将(jiang)该片(pian)(pian)段(duan)注(zhu)射到NPG小(xiao)鼠的(de)原核中(zhong)。在获得的(de)后(hou)代中(zhong)筛选到合适表(biao)达量的(de)阳性(xing)小(xiao)鼠。


2. 表型

在NPG小(xiao)鼠(shu)(shu)中表(biao)达(da)人(ren)(ren)的(de)(de)hCSF2细(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)因(yin)子(zi)。CSF2,又称为GM-CSF(粒细(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)-巨(ju)(ju)噬(shi)细(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)集落刺激(ji)(ji)因(yin)子(zi)),是(shi)一种造血生长(zhang)因(yin)子(zi),刺激(ji)(ji)中性(xing)(xing)粒细(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)和(he)(he)(he)(he)巨(ju)(ju)噬(shi)细(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)的(de)(de)发育(yu),促进早期红系(xi)(xi)巨(ju)(ju)核细(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)和(he)(he)(he)(he)嗜酸性(xing)(xing)祖细(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)的(de)(de)增(zeng)殖(zhi)和(he)(he)(he)(he)发育(yu)。hCSF2 NPG小(xiao)鼠(shu)(shu),表(biao)达(da)合适(shi)含量的(de)(de)人(ren)(ren)CSF2蛋白,与表(biao)达(da)人(ren)(ren)M-CSF和(he)(he)(he)(he)IL3细(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)因(yin)子(zi)的(de)(de)NPG小(xiao)鼠(shu)(shu)一起,在移植人(ren)(ren)CD34+造血干细(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)6-8周后(hou),于外周血、骨髓、胸(xiong)腺和(he)(he)(he)(he)脾脏(zang)以(yi)及(ji)包括肺和(he)(he)(he)(he)肝(gan)在内的(de)(de)非淋(lin)巴(ba)组织(zhi)中,形成稳(wen)定广泛的(de)(de)髓系(xi)(xi)和(he)(he)(he)(he)淋(lin)巴(ba)系(xi)(xi)细(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)分化(hua)。在血液和(he)(he)(he)(he)组织(zhi)中,可检测到粒细(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)分化(hua)(嗜碱(jian)性(xing)(xing)粒细(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)、中性(xing)(xing)粒细(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)和(he)(he)(he)(he)肥大(da)细(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)),抗原(yuan)呈(cheng)递细(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)分化(hua)(树突状细(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)和(he)(he)(he)(he)巨(ju)(ju)噬(shi)细(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao))和(he)(he)(he)(he)调节性(xing)(xing)T细(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)群体等。表(biao)达(da)人(ren)(ren)M-CSF、GM-CSF和(he)(he)(he)(he)人(ren)(ren)IL-3细(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)因(yin)子(zi)与Hu-NPG人(ren)(ren)源化(hua)小(xiao)鼠(shu)(shu)相比,人(ren)(ren)类(lei)(lei)造血干细(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)(hsc)的(de)(de)整体植入水平更高,分化(hua)细(xi)(xi)(xi)(xi)(xi)(xi)(xi)胞(bao)(bao)(bao)种类(lei)(lei)更多。


3. 应用领域

-人源化和癌症(zheng)治疗

-人体过敏反应模型,免疫、造血移植重建模型

-感染性(xing)疾病

-再生医学


4. 参(can)考文献

(1)Ito R, Takahashi T, Katano I, Kawai K, Kamisako T, Ogura T, Ida-Tanaka M, Suemizu H, Nunomura S, Ra C, Mori A, Aiso S, Ito M. (2013) Establishment of a human allergy model using human IL-3/GM-CSF-transgenic NOG mice. J Immunol.191(6):2890-9.


(2)Wunderlich M; Chou FS; Link KA; Mizukawa B; Perry RL; Carroll M; Mulloy JC. 2010. AML xenograft efficiency is significantly improved in NOD/SCID-IL2RG mice constitutively expressing human SCF, GM-CSF and IL-3. Leukemia 24(10):1785-8


(3)Rongvaux A; Willinger T; Martinek J; Strowig T; Gearty SV; Teichmann LL; Saito Y; Marches F; Halene S; Palucka AK; Manz MG; Flavell RA. 2014. Development and function of human innate immune cells in a humanized mouse model. Nat Biotechnol 32(4):364-72