簡要描述：TAKARA T細胞擴增GT-T551 H3無血清培養(yǎng)基：
GT-T551培養(yǎng)基支持Anti-CD3/IL-2介導(dǎo)的T細胞刺激活化。用這種方法激活T細胞時，如添加RetroNectin重組人纖維蛋白片段可以增強T細胞的擴增，而且所得T細胞比單獨使用CD3單抗產(chǎn)生更多 naïve T cells（初始T細胞）。

T細胞擴增GT-T551 H3無血清培養(yǎng)基

產(chǎn)品英文名稱：GT-T551 H3 Culture medium, 1L Bottle

產(chǎn)品中文名稱：GT-T551 H3無血清培養(yǎng)基

適用：淋巴細胞、DC細胞、NK細胞培養(yǎng)

特別應(yīng)用：T細胞擴增，與CD3單抗一起活化T細胞

保存：2℃ to 8℃

產(chǎn)品形態(tài)：透明液體

組份：GT-T551 Culture medium, 1L Bottle  1,000 mL

GT-T551培養(yǎng)基適合人T淋巴細胞（T細胞）的擴增。培養(yǎng)基添加了人白蛋白（醫(yī)藥級別），重組人胰島素，L-谷氨酰胺和鏈霉素，不含其它蛋白和生長因子。不論是否添加血清，GT-T551都能支持細胞穩(wěn)定生長。

GT-T551培養(yǎng)基支持Anti-CD3/IL-2介導(dǎo)的T細胞刺激活化。用這種方法激活T細胞時，如添加RetroNectin重組人纖維蛋白片段可以增強T細胞的擴增，而且所得T細胞比單獨使用CD3單抗產(chǎn)生更多 naïve T cells（初始T細胞）。

GT-T551 培養(yǎng)基產(chǎn)品特點

日本*無血清培養(yǎng)基，已經(jīng)加入人血白蛋白組分。

支持淋巴細胞的高密度細胞培養(yǎng)。

在日本多家醫(yī)院用于臨床治療的細胞培養(yǎng)。

配制用水符合國家藥監(jiān)局《人體細胞治療研究和制劑質(zhì)量控制指導(dǎo)原則》中注射用水標(biāo)準(zhǔn)。

GT-T551 培養(yǎng)液在使用前加入病人0.6%自體血漿，性能可以和許多高價培養(yǎng)液效果相同。

GT-T551 H3 是GT-T551的升級版培養(yǎng)基，采用全新的緩沖系統(tǒng)，可用于無血漿培養(yǎng)，CD3+CD56+比例高。如果培養(yǎng)時添加病人自體血漿，效果更為顯著。

GT-T551培養(yǎng)基性能

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.

細胞擴增速度

人PBMCs（外周血單個核細胞）經(jīng)CD3單抗刺激活化4天后，使用GT-T551培養(yǎng)基擴增培養(yǎng)。相比其他培養(yǎng)基，GT-T551在沒有添加人AB血清的情況下仍然能保持T細胞穩(wěn)定生長。

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.

培養(yǎng)體系：GT-T551培養(yǎng)基，CD3單抗，CultiLife培養(yǎng)袋，添加0.6%的熱滅活血漿。添加RetroNectin重組人纖維蛋白片段后，T細胞擴增大幅上升，并且得到高比例的初始T細胞。

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

GT-T551和CultiLife 培養(yǎng)袋一起使用后，T細胞擴增效率更高。

使用GT-T551培養(yǎng)基進行T細胞擴增流程

培養(yǎng)袋預(yù)包被抗CD3單克隆抗體和RetroNectin重組人纖維蛋白片段。

加入PBMCs。

第四天傳代。

收獲細胞。

T細胞擴增方法詳見TAKARA GT-T551說明書。

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.

參考文獻

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.

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