Brown R L, Ortaldo J R, Griffith R L, Blanca I, Rabin H
J Immunol Methods. 1985 Aug 2;81(2):207-14. doi: 10.1016/0022-1759(85)90205-4.
We recently developed a serum-free (SF) culture medium that supports the growth of several established lymphoid cell lines. In an effort to develop a standardized medium for assay of human natural killer (NK) cell activity, we compared the cytotoxic activity of peripheral blood mononuclear leukocytes (PBL) and purified large granular lymphocytes (LGL) cultured in SF medium containing interleukin 2 (IL-2) or medium containing 10% fetal bovine serum (FBS) plus IL-2. The results indicated that PBL had a 30% increase in cumulative net cell growth and had as high or higher cytotoxic activity after growth in SF medium than in medium containing FBS. Purified LGL had a 50% increase in cumulative net cell growth and persisted approximately 2 weeks longer in culture in medium containing FBS than in SF medium. However, the cytotoxic activity of cells grown in SF medium persisted during the initial 3 weeks of culture. Purified LGL that were maintained and were subcultured at cell densities of 10(6) cells or greater per milliliter of either SF or FBS-containing medium had equivalent levels of cytotoxicity over a 44-day period in either medium compared with cells subcultured at a density of 5 X 10(5) cells per milliliter of medium. NK cells produced a cytotoxic factor (NKCF) in SF medium, and its cytotoxic activity was blocked by 10% FBS. We conclude that the SF medium supplemented with IL-2 can be used as an alternative to FBS-containing medium with IL-2 for the growth of NK cells and is advantageous for the production of NKCF.
我们最近开发了一种无血清（SF）培养基，可支持多种已建立的淋巴细胞系生长。为了开发一种用于检测人类自然杀伤（NK）细胞活性的标准化培养基，我们比较了在含有白细胞介素2（IL-2）的SF培养基或含有10%胎牛血清（FBS）加IL-2的培养基中培养的外周血单个核白细胞（PBL）和纯化的大颗粒淋巴细胞（LGL）的细胞毒性活性。结果表明，PBL的累积净细胞生长增加了30%，并且在SF培养基中生长后的细胞毒性活性与在含有FBS的培养基中生长后的细胞毒性活性一样高或更高。纯化的LGL的累积净细胞生长增加了50%，并且在含有FBS的培养基中培养的时间比在SF培养基中大约长2周。然而，在SF培养基中生长的细胞的细胞毒性活性在培养的最初3周内持续存在。与以每毫升培养基5×10⁵个细胞的密度传代培养的细胞相比，在每毫升SF或含FBS的培养基中以10⁶个细胞或更高的细胞密度维持并传代培养的纯化LGL在44天的时间内具有同等水平的细胞毒性。NK细胞在SF培养基中产生一种细胞毒性因子（NKCF），其细胞毒性活性被10% FBS阻断。我们得出结论，补充IL-2的SF培养基可作为含IL-2的含FBS培养基的替代品用于NK细胞的生长，并且有利于NKCF的产生。
