發(fā)布時間：2018-05-03 23:18

  本文選題：細(xì)胞因子誘導(dǎo)的殺傷細(xì)胞 + 培養(yǎng)基��； 參考：《中國腫瘤生物治療雜志》2014年06期

【摘要】：目的:探討不同細(xì)胞培養(yǎng)基及血清的培養(yǎng)體系對細(xì)胞因子誘導(dǎo)殺傷(cytokine-induced killer,CIK)細(xì)胞的增殖和功能的影響。方法:采集6例肺癌患者外周血,分離獲得單個核細(xì)胞,按照不同血清與培養(yǎng)基分為8組:GT-T551培養(yǎng)基+患者自體血清組、GT-T551培養(yǎng)基+健康人血清組、GT-T551培養(yǎng)基+FBS組、GT-T551培養(yǎng)基組、RPMI 1640培養(yǎng)基+患者自體血清組、RPMI 1640培養(yǎng)基+健康人血清組、RPMI 1640培養(yǎng)基+FBS組及RPMI 1640培養(yǎng)基組。采用CFSE染色法檢測細(xì)胞增殖能力,流式細(xì)胞術(shù)檢測CIK細(xì)胞CD3+CD8+T細(xì)胞及CD3+CD4+T細(xì)胞顆粒酶B、IFN-γ、穿孔素的分泌情況及以白血病NB4、K562細(xì)胞為靶細(xì)胞時CD3+CD56+T細(xì)胞、CD3+CD4+T細(xì)胞及CD3+CD8+T細(xì)胞表面CD107a的表達(dá)。結(jié)果:GTT551培養(yǎng)基加入患者自體血清組CIK細(xì)胞的增殖指數(shù)顯著高于其他各組(P0.05)。GT-T551培養(yǎng)基或RPMI 1640培養(yǎng)基中加入患者自體血清組CD4+T細(xì)胞顆粒酶B的分泌水平均顯著高于加健康人血清組[(22.85±3.50)%vs(13.28±1.75)%,(22.57±3.45)%vs(15.37±4.08)%,均P0.01],而且GT-T551+患者自體血清組CD8+T細(xì)胞分泌IFN-γ的能力顯著高于加健康人血清組(P0.05)。以白血病細(xì)胞系NB4和K562細(xì)胞作為靶細(xì)胞時,檢測CD3+CD56+NKT細(xì)胞表面CD107a的表達(dá),GT-T551培養(yǎng)基中加入患者自體血清組優(yōu)于加健康人血清組[(7.10±1.94)%vs(2.73±0.79)%,(8.00±1.82)%vs(3.03±0.78)%,P0.01],同時優(yōu)于RPMI1640+患者自體血清組[(4.45±1.96)%、(3.30±1.47)%,P0.01]。結(jié)論:GT-T551培養(yǎng)基加患者自體血清的培養(yǎng)體系更有利于CIK細(xì)胞的增殖、細(xì)胞因子分泌及發(fā)揮殺傷功能,可推薦作為CIK細(xì)胞最佳培養(yǎng)體系。
[Abstract]:Aim: to investigate the effects of different cell culture medium and serum on the proliferation and function of cytokine-induced killer CIK cells. Methods: mononuclear cells were isolated from peripheral blood of 6 patients with lung cancer. According to different serum and medium, 8 groups were divided into 8 groups: GT-T551, patient's autoserum group, healthy human serum group, FBS medium, GT-T551 medium, patient's autoserum group, RPMI1640 medium, healthy medium. Human serum group: RPMI#number0# medium FBS group and RPMI 1640 medium group. Cell proliferation was detected by CFSE staining. Flow cytometry was used to detect the secretion of CD3 CD8 T cells and CD3 CD4 T cells granzyme Bn- 緯, perforin, and the expression of CD107a on CD3 CD56 T cells, CD3 CD4 T cells and CD3 CD8 T cells. Results the proliferative index of CIK cells in the patients with autologous serum was significantly higher than that in the other groups (P 0.05N. GT-T551 or RPMI 1640). The secretory levels of granzyme B of CD4 T cells were significantly higher in the patients with autologous serum addition than those in the other groups. The ability of CD8 T cells to secrete IFN- 緯 in autologous serum of patients with GT-T551 was significantly higher than that in the serum of healthy persons (22.85 鹵1.75 鹵22.57 鹵3.45)%vs(15.37 鹵4.08, P0.01), and the ability of secreting IFN- 緯 by CD8 T cells in autologous serum of GT-T551 patients was significantly higher than that in the control group (P 0.05). When the leukemic cell lines NB4 and K562 cells were used as target cells, the expression of CD107a on the surface of CD3 CD56 NKT cells was detected in GT-T551 medium. The addition of autologous serum from patients was superior to that of healthy persons [7.10 鹵1.94)%vs(2.73 鹵0.79g + 0.78P 0.01], and it was also better than that of RPMI1640 patients [4.45 鹵1.960.30 鹵1.47P 0.01]. Conclusion the cell proliferation, cytokine secretion and killing function of CIK cells are more favorable to the culture system of the cell culture medium of: GT-T551 and patient's autologous serum, and can be recommended as the best culture system for CIK cells.
【作者單位】： 鄭州大學(xué)第一附屬醫(yī)院生物細(xì)胞治療中心;鄭州大學(xué)第一附屬醫(yī)院腫瘤科;鄭州大學(xué)生命科學(xué)院;鄭州大學(xué)第一附屬醫(yī)院消化內(nèi)科;河南省高等學(xué)校臨床醫(yī)學(xué)重點學(xué)科開放實驗室;
【基金】：國家自然科學(xué)基金資助項目(No.81171986,No.81271815) 衛(wèi)生部科研攻關(guān)基金資助(No.20110110001) 河南省科技廳基礎(chǔ)與前沿技術(shù)研究基金資助(No.112300410153,No.122300410155) 河南省科技廳科技創(chuàng)新人才計劃資助(No.124200510006) 鄭州大學(xué)第一附屬醫(yī)院院內(nèi)創(chuàng)新團(tuán)隊基金資助~~
【分類號】：R329.2

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