Long-term human peripheral blood monocyte cultures: establishment, metabolism and morphology of primary human monocyte-macrophage cell cultures.
Long-term human peripheral blood monocyte cultures: establishment, metabolism and morphology of primary human monocyte-macrophage cell cultures.
复制标题
长期人外周血单核细胞培养物：原代人单核细胞-巨噬细胞培养物的建立、代谢和形态。
作者：
S. Zuckerman;S. Ackerman;S. Douglas
人外周血单核细胞在体外培养中使用非人血清源维持长达4个月。单核细胞在Dulbecco改良Eagle培养基中培养，用20 mM HEPES缓冲，含有10%马血清和10%胎牛血清。采用微滴法、Linbro法和t25法进行体外代谢和形态变化的研究。在培养过程中，单核细胞的大小增加，扫描电镜显示其膜活性增加，并分化成纺锤状和上皮样形态的形态学异质性群体。这些细胞类型保留了吞噬E过剩和EA以及与EA和EAC结簇的能力。在培养过程中也观察到较大的巨多核细胞；其中许多缺乏结合或吞噬惰性红细胞或抗体包被红细胞的能力。在培养过程中，溶菌酶释放量和酸性磷酸酶活性也有所增加。培养的单核细胞表现出亮氨酸和尿苷摄取的特征，培养5天后观察到最大活性。没有检测到胸腺嘧啶的摄取。该体外系统可对涉及单核细胞生长和分化的调控过程进行详细分析。
Human peripheral blood monocytes were maintained in in vitro culture for periods up to 4 months using a non-human serum source. Monocytes were cultured in Dulbecco's modified Eagle's medium buffered with 20 mM HEPES and containing 10% horse serum and 10% foetal calf serum. The metabolic and morphological changes which occur in vitro were investigated using microtitre, Linbro and T 25 cultures. During culture, monocytes increased in size, had increased membrane activity as visualized by SEM, and differentiated into a morphologically heterogeneous population of fusiform and epithelioid shapes. These cell types retained the ability to phagocytose E glut and EA and to rosette with EA and EAC. Larger giant polynucleated cells were also observed during culture; many of these lacked the ability to bind or phagocytose inert or antibody-coated erythrocytes. Increases in lysozyme release and acid phosphatase activity also occurred during culture. Cultured monocytes exhibited characteristic profiles of leucine and uridine uptake with maximal activity observed by 5 days of culture. There was no detectable uptake of thymidine. Detailed analysis of regulatory processes involved in monocyte growth and differentiation could be performed with this in vitro system.