GT-T551培养基适合人T淋巴细胞（T细胞）的扩增。培养基添加了人白蛋白（医药级别），重组人胰岛素，L-谷氨酰胺和链霉素，不含其它蛋白和生长因子。不论是否添加血清，GT-T551都能支持细胞稳定生长。

GT-T551培养基支持Anti-CD3/IL-2介导的T细胞刺激活化。用这种方法激活T细胞时，如添加RetroNectin重组人纤维蛋白片段可以增强T细胞的扩增，而且所得T细胞比单独使用CD3单抗产生更多 naïve T cells（初始T细胞）。

GT-T551 培养基产品特点

日本原装进口无血清培养基，已经加入人血白蛋白组分。

支持淋巴细胞的高密度细胞培养。

在日本多家医院用于临床治疗的细胞培养。

配制用水符合国家药监局《人体细胞治疗研究和制剂质量控制指导原则》中注射用水标准。

GT-T551 培养液在使用前加入病人0.6%自体血浆，性能可以和许多高价培养液效果相同。

GT-T551 H3 是GT-T551的升级版培养基，采用全新的缓冲系统，可用于无血浆培养，CD3+CD56+比例高。如果培养时添加病人自体血浆，效果更为显著。

GT-T551培养基性能

细胞扩增速度 

Human PBMCs were cultured for 4 days with anti-CD3 stimulation and further cultured for 10 days with GT-T551 culture medium or competitors’ media products. GT-T551 showed stable expansion of T cells even without the addition of human serum.

人PBMCs（外周血单个核细胞）经CD3单抗刺激活化4天后，使用GT-T551培养基扩增培养。相比其他培养基，GT-T551在没有添加人AB血清的情况下仍然能保持T细胞稳定生长。

高比例初始T细胞

PBMCs and autologous plasma were prepared using 50 ml of blood from two healthy volunteers on the basis of informed consent. The effect of RetroNectin (RN) co-stimulation on T-cell expansion was tested using a culture system consisting of GT-T551 Culture medium, IL-2, anti-CD3 mAb, and CultiLife Culture bags, with or without RN. Heat-inactivated plasma at a final concentration of 0.6% was added to the GT-T551 Culture medium at Day 0 and Day 4. RN co-stimulation resulted in a dramatic increase in T-cell expansion and the population contained a high proportion of naïve T cells.

培养体系：GT-T551培养基，CD3单抗，CultiLife培养袋，添加0.6%的热灭活血浆。添加RetroNectin重组人纤维蛋白片段后，T细胞扩增大幅上升，并且得到高比例的初始T细胞。

GT-T551 is effective at expanding T lymphocytes in combination with CultiLife culture bags, as seen here.

GT-T551和CultiLife 培养袋一起使用后，T细胞扩增效率更高。

 使用GT-T551培养基进行T细胞扩增流程

培养袋预包被抗CD3单克隆抗体和RetroNectin重组人纤维蛋白片段。

加入PBMCs。

第四天传代。

收获细胞。

T细胞扩增方法详见TAKARA GT-T551说明书。

GT-T551 is a culture medium ideally suited for expansion of human T lymphocytes (T cells). It is supplemented with human albumin (pharmaceutical grade), recombinant human insulin, L-glutamine, and streptomycin; no additional proteins or growth factors are added. The medium can support stable growth in the presence or absence of serum.

Anti-CD3/IL-2 T-cell activation can also be performed in this medium (Wang et al. 2013; Ai et al. 2014; Dodo et al. 2014). When using this activation method, the additional presence of RetroNectin enhances the proliferation of T cells (Yu et al. 2008; Ishikawa et al. 2014; Hosoi et al. 2014; Sakamoto et al. 2015; Li et al. 2015) and results in an expanded cell population that shows a higher proportion of naïve T cells than anti-CD3 antibody stimulation alone (Hosoi et al. 2014).

Features

Used for extended culture or expansion of T cells

Supplemented with human serum albumin, human insulin, L-glutamine, and streptomycin

Endotoxin tested (endotoxin <0.3 EU/ml)

Compatible with CultiLife Culture Bags for large-scale expansion of T cells

Anti-CD3/IL-2 T-cell activation in the presence of RetroNectin enhances proliferation and leads to a higher proportion of naïve T cells. For more information, the user manual can be found under the Documents tab.

The method of T-cell expansion by RetroNectin stimulation requires a license from Takara Bio Inc. for uses other than research.

参考文献

1)  Wang, Y.,et al. (2013) CIK cells from recurrent or refractory AML patients can be efficiently expanded in vitro and used for reduction of leukemic blasts in vivo.Exp Hematol. 41(3): 241-252.

2)  Ai, Y. Q.,et al. (2014) The clinical effects of dendritic cell vaccines combined with cytokine-induced killer cells intraperitoneal injected on patients with malignant ascites. Int J Clin Exp Med. 7(11): 4272-4281.

3)  Dodo, K.et al. (2014) An efficient large-scale retroviral transduction method involving preloading the vector into a RetroNectin-coated bag with low-temperature shaking.PLoS ONE. 9(1): e86275.

4)  Yu, S. S,et al. (2008) In vivo persistence of genetically modified T cells generated ex vivo using the fibronectin CH296 stimulation method.Cancer Gene Ther. 15(8):508-516.

5)  Ishikawa, T.,et al. (2014) Phase I clinical trial of fibronectin CH296-stimulated T cell therapy in patients with advanced cancer.PLoS ONE. 9(1): e83786.

6)  Hosoi, H.,et al. (2014) Stimulation through very late antigen-4 and -5 improves the multifunctionality and memory formation of CD8+ T cells.Eur J Immunol. 44(6):1747-1758.

7)  Sakamoto, N.,et al. (2015) Phase I clinical trial of autologous NK cell therapy using novel expansion method in patients with advanced digestive cancer.J Transl Med.13: 277.

8)  Li, W.,et al. (2015) Efficacy of RetroNectin-activated cytokine-induced killer cell therapy in metastatic brain tumor patients.Oncol Res Treat. 38(4): 160-165.