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  • 1
    ISSN: 1520-6904
    Source: ACS Legacy Archives
    Topics: Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Cytotechnology 6 (1991), S. 33-38 
    ISSN: 1573-0778
    Keywords: hybridoma ; monoclonal antibody ; myeloma ; serum-free and tissue plasminogen activator
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract We established a serum-free medium of low protein content(125μg/ml) TYI 100, consisting of three hormones and five growth factors for the growth of lymphoid and hybridoma cell lines. In TYI 100 medium, mouse and human hybridomas grew equally well as in RPMI 1640 supplemented with 10% fetal bovine serum (10% FBS) without adaptation to the serum-free medium. TYI 100 medium allowed several passages of mouse hybridoma lines and the total cell number was more than in 10% FBS. TYI 100 medium also supported growth of myelomas and anchorage dependent cell lines, Bowes and CHO, well. TYI 100 medium is composed of inexpensive supplements and is therefor applicable to large scale culture.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1573-0778
    Keywords: cell culture ; lymphocyte ; lymphokine-activated killer cell ; recombinant interleukin 2 ; serum-free medium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract We examined a serum-free medium (designated as TYI 101) for the generation of lymphokine-activated killer (LAK) cells from human lymphocytes, regional lymph node lymphocytes (RLNL) and peripheral blood lymphocytes (PBL). TYI 101 medium consisted of, in addition to nutrient mixture, transferrin, insulin, fetuin, sodium selenite, 2-mercaptoethanol, o-phosphorylethanolamine, chick egg yolk and porcine kidney extract. These hormones were effective for supporting RLNL proliferation as assessed by (3H)-thymidine uptake. When human lymphocytes from two different sources were cultivated with recombinant interleukin 2 (rIL-2) in TYI 101 medium, LAK activity was generated. In cultures of PBL from a healthy donor, LAK cells were generated in TYI 101 medium as efficiently as in RPMI 1640 medium supplemented with 10% human AB-type serum (RPMI-AB). In cultures of RLNL from lung cancer patients, LAK activity obtained in TYI 101 medium was about sixty-five percent of that in RPMI-AB. However, the addition of a small amount of AB-type serum improved the generation of LAK activity, LAK cell expansion, and cell viability in TYI 101 medium. We conclude that TYI 101 medium can be used for the generation of LAK cells from human lymph node lymphocytes with supplementation of none or only a reduced amount of human serum.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1573-0778
    Keywords: adoptive immunotherapy ; cell culture ; cell culture apparatus ; Interleukin-2 ; lymphokine-activated killer cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: 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 2.7×107 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. The JCC-device was able to reduce the consumption of basal medium, rIL-2 and serum by 20%, 84% and 96%, respectively compared to the conventional tissue culture systems. The JCC-device improved the routine performance of adoptive immunotherapy with LAK cells and rIL-2.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1573-0778
    Keywords: cell culture ; cell culture apparatus ; dialysis membrane ; perfusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract We recently developed a new dialysis culture system (termed LIFROC-device) for the cultivation of lymphokine-activated killer cells (Murataet al., 1990, 1991). In the present study, we applied the LIFROC-device (400 ml culture vessel) to the cultivation of mammalian cells for the production of biologically active substances. We cultured mouse-mouse hybridoma TP-709, secreting anti-tissue plasminogen activator (tPA) monoclonal antibody (mAb), recombinant CHO GT19, secreting hGH, and human melanoma Bowes cells, secreting tPA. With the LIFROC-device, TP-709 grew to a maximal cell density of 3.8×106 cells/ml and and produced 480 μg/ml (192 mg in total) of mAb. GT19 reached a cell density of 2.2×106 cells/ml and produced 302 μg/ml (120 mg in total) of hGH. Bowes cells expanded to 4.4×106 cells/ml and secreted 8.5 μg/ml (3.3 mg in total) of tPA. The protein concentration in the culture broths of the LIFROC-device became 7–200 times higher than that of batch culture. Thus, the LIFROC-device can be applied to protein production as well as cell growth with high efficiency.
    Type of Medium: Electronic Resource
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