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人淋巴因子激活的杀伤细胞新培养系统的开发:与传统静态培养方法的比较。

Development of a new culture system for human lymphokine-activated killer cells: comparison with a conventional static culture method.

作者信息

Murata M, Yano T, Yoshino I, Togami M, Sogabe M, Yasumoto K, Sugimachi K, Kimura G, Nomoto K

机构信息

Department of Immunology, Kyushu University, Fukuoka, Japan.

出版信息

Cytotechnology. 1991 Oct;7(2):75-83. doi: 10.1007/BF00350913.

DOI:10.1007/BF00350913
PMID:1367909
Abstract

We recently developed a new culture system based on dialysis perfusion (designated JCC-device) for the generation and expansion of human lymphokine-activated killer (LAK) cells (Murata et al., 1990). More recently we have scaled up the volume of the culture vessel of the JCC-device from 100 ml to 400 ml for clinical use. In the present study, using this new 400 ml JCC-device, we cultured human lymph node lymphocytes (LNL) obtained from 8 surgical patients with primary lung cancer, and investigated the cellular characteristics in comparison with a conventional batchwise culture system using tissue culture dishes. With the JCC-device, the cell density reached a maximum 2.7 x 10(7) cells/ml with greater than 90% viability by the appropriate exchange of perfusion medium and by making additions at the appropriate intervals for recombinant interleukin-2 (rIL-2). The expansion fold of LNL with the JCC-device, ranging 6.6- to 19.2-fold (mean 13.8-fold), was not significantly different from that in dish cultures. There was no marked difference in cell surface phenotypes between the two culture systems in 7 out of 8 cases. As for LAK activity of LNL, the JCC culture was either superior or equal in 4 out of 8 cases, but inferior in the other 4 cases to the conventional dish cultures. In the latter cases, the usage of serum for the JCC culture was limited, which might have resulted in the low LAK activity.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

我们最近开发了一种基于透析灌注的新培养系统(称为JCC装置),用于人淋巴因子激活的杀伤细胞(LAK细胞)的生成和扩增(村田等人,1990年)。最近,我们已将JCC装置的培养容器体积从100毫升扩大到400毫升以供临床使用。在本研究中,我们使用这种新的400毫升JCC装置,培养了从8名原发性肺癌手术患者获得的人淋巴结淋巴细胞(LNL),并与使用组织培养皿的传统分批培养系统相比,研究了细胞特性。使用JCC装置,通过适当更换灌注培养基并在适当间隔添加重组白细胞介素-2(rIL-2),细胞密度最高达到2.7×10⁷个细胞/毫升,活力大于90%。JCC装置培养的LNL扩增倍数在6.6至19.2倍之间(平均13.8倍),与培养皿培养的扩增倍数无显著差异。8例中有7例在两种培养系统之间的细胞表面表型无明显差异。至于LNL的LAK活性,8例中有4例JCC培养优于或等同于传统培养皿培养,但在其他4例中低于传统培养皿培养。在后者的情况下,JCC培养中血清的使用受限,这可能导致了低LAK活性。(摘要截短至250字)

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Cytotechnology. 1991 Oct;7(2):75-83. doi: 10.1007/BF00350913.
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引用本文的文献

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A new culture system for the cultivation of mammalian cells for the production of several biologically active substances.一种用于培养哺乳动物细胞以生产多种生物活性物质的新培养系统。
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本文引用的文献

1
Cytotoxicity of a factor isolated from human spleen.从人脾脏中分离出的一种因子的细胞毒性
J Natl Cancer Inst. 1973 Feb;50(2):535-8. doi: 10.1093/jnci/50.2.535.
2
Morphological and virological investigations on cultured Burkitt tumor lymphoblasts (strain Raji).对培养的伯基特淋巴瘤淋巴母细胞(拉吉株)进行的形态学和病毒学研究。
J Natl Cancer Inst. 1966 Oct;37(4):547-59.
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Purification and characterization of recombinant human interleukin-2 produced in Escherichia coli.在大肠杆菌中产生的重组人白细胞介素-2的纯化及特性鉴定
Biochem Biophys Res Commun. 1985 Jul 31;130(2):692-9. doi: 10.1016/0006-291x(85)90472-3.
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Constant-infusion recombinant interleukin-2 in adoptive immunotherapy of advanced cancer.持续输注重组白细胞介素-2在晚期癌症过继性免疫治疗中的应用
N Engl J Med. 1987 Apr 9;316(15):898-905. doi: 10.1056/NEJM198704093161502.
5
A progress report on the treatment of 157 patients with advanced cancer using lymphokine-activated killer cells and interleukin-2 or high-dose interleukin-2 alone.关于使用淋巴因子激活的杀伤细胞和白细胞介素-2或单独使用高剂量白细胞介素-2治疗157例晚期癌症患者的进展报告。
N Engl J Med. 1987 Apr 9;316(15):889-97. doi: 10.1056/NEJM198704093161501.
6
Phenotypic characterization of lymphokine-activated killer cells from human lymph node lymphocytes.人淋巴结淋巴细胞来源的淋巴因子激活的杀伤细胞的表型特征
Cell Immunol. 1989 Sep;122(2):578-84. doi: 10.1016/0008-8749(89)90104-4.
7
Generation and expansion of lymphokine-activated killer cells from lymph node lymphocytes in human lung cancer.从人肺癌患者的淋巴结淋巴细胞中生成和扩增淋巴因子激活的杀伤细胞
Eur J Cancer Clin Oncol. 1989 Feb;25(2):201-8. doi: 10.1016/0277-5379(89)90009-6.
8
Long-term growth of lymphokine-activated killer (LAK) cells: role of anti-CD3, beta-IL 1, interferon-gamma and -beta.淋巴因子激活的杀伤细胞(LAK)的长期生长:抗CD3、β-白细胞介素-1、干扰素-γ和-β的作用
J Immunol. 1987 Apr 15;138(8):2728-33.
9
Characterization and modulation of human lymphokine (interleukin 2) activated killer cell induction.人淋巴因子(白细胞介素2)激活的杀伤细胞诱导的特性与调控
Cancer Res. 1986 Jun;46(6):2834-8.
10
Development of a new culture system for human lymphokine-activated killer cells.一种用于人淋巴因子激活杀伤细胞的新型培养系统的研发。
J Immunol Methods. 1990 May 8;129(1):89-95. doi: 10.1016/0022-1759(90)90424-t.