Vincent F, Eischen A, Bergerat J P, Faradji A, Bohbot A, Oberling F
Institute for Hematology and Immunology, Faculty of Medicine, Louis Pasteur University, Strasbourg, France.
Exp Hematol. 1992 Jan;20(1):17-23.
Large quantities of human blood-derived monocytes have been cultured in suspension in nonadherent cell culture bags and maintained for up to 3 weeks in a serum-free medium. This serum-free medium contained Iscove's modified Dulbecco's medium (IMDM) supplemented with human albumin, alpha-phosphatidylcholine, transferrin, and insulin. Morphology, cell surface antigens, and functional properties of these in vitro maturing macrophages were studied in comparison with macrophages cultured in a standard medium containing 10% fetal calf serum. In this report we demonstrate that this serum-free medium allows a better yield of cell survival than the standard medium; it also allows the differentiation of blood monocytes into fully functional macrophagic cells that express the different antigens found in mature macrophages. The results indicate that the use of serum-free defined medium offers good conditions in which to culture large numbers of human monocytes and allows an accurate analysis of the effect of supplementation with growth factors such as granulocyte-macrophage colony-stimulating factor (GM-CSF) and macrophage colony-stimulating factor (M-CSF) on the differentiation and survival of monocytes and macrophages. Serum-free cultures could also be helpful for the precise analysis of the cell secretion activity and for determining the factors that are responsible for monocyte maturation into macrophages.
大量源自人类血液的单核细胞已在非贴壁细胞培养袋中进行悬浮培养，并在无血清培养基中维持长达3周。这种无血清培养基含有伊思柯夫改良的杜尔贝科培养基（IMDM），补充了人白蛋白、α-磷脂酰胆碱、转铁蛋白和胰岛素。将这些体外成熟巨噬细胞的形态、细胞表面抗原和功能特性与在含有10%胎牛血清的标准培养基中培养的巨噬细胞进行了比较研究。在本报告中，我们证明这种无血清培养基比标准培养基能产生更高的细胞存活率；它还能使血液单核细胞分化为功能完全的巨噬细胞，这些巨噬细胞表达成熟巨噬细胞中发现的不同抗原。结果表明，使用无血清限定培养基为培养大量人类单核细胞提供了良好条件，并能准确分析补充粒细胞-巨噬细胞集落刺激因子（GM-CSF）和巨噬细胞集落刺激因子（M-CSF）等生长因子对单核细胞和巨噬细胞分化及存活的影响。无血清培养对于精确分析细胞分泌活性以及确定单核细胞成熟为巨噬细胞的相关因子也可能有帮助。
