Cushion M T, Ruffolo J J, Linke M J, Walzer P D
Exp Parasitol. 1985 Aug;60(1):43-54. doi: 10.1016/s0014-4894(85)80021-7.
Recent studies indicate that rat Pneumocystis carinii can be propagated in the A549 cell line, an alveolar epithelioid cell line derived from human lung carcinoma. In the present study, growth of P. carinii was compared in the A549 cell line and the WI-38 VA13 subline 2RA, an SV40 transformed derivative of the human fetal fibroblast cell line with epithelioid morphology. Similar P. carinii growth occurred in both cell lines under optimal conditions, but the WI-38 VA13 cell line was usually more sensitive to changes in the culture system. Growth of P. carinii was affected by temperature, environmental gas mixture, motion of the cultures, and source and concentration of serum additives, but not by the presence of antibodies in the medium. A technique was developed for quantitating P. carinii in the lung inoculum which permitted analysis of P. carinii growth during the first 24 hr of culture. Inverted microscope and oil immersion phase-contrast microscopy were very helpful in monitoring the organism's stages of development and viability. Thus, this culture system should be helpful in establishing standard methodology for in vitro work with P. carinii.
近期研究表明，大鼠卡氏肺孢子虫可在A549细胞系中繁殖，A549细胞系是一种源自人肺癌的肺泡上皮样细胞系。在本研究中，比较了卡氏肺孢子虫在A549细胞系和WI-38 VA13亚系2RA中的生长情况，WI-38 VA13亚系2RA是具有上皮样形态的人胎儿成纤维细胞系经SV40转化后的衍生物。在最佳条件下，两种细胞系中卡氏肺孢子虫的生长相似，但WI-38 VA13细胞系通常对培养系统的变化更敏感。卡氏肺孢子虫的生长受温度、环境气体混合物、培养物的运动以及血清添加剂的来源和浓度影响，但不受培养基中抗体存在的影响。开发了一种定量肺接种物中卡氏肺孢子虫的技术，该技术可用于分析培养最初24小时内卡氏肺孢子虫的生长情况。倒置显微镜和油浸相差显微镜对于监测该生物体的发育阶段和活力非常有帮助。因此，这种培养系统应有助于建立卡氏肺孢子虫体外研究的标准方法。
