Pneumocystis carinii: growth variables and estimates in the A549 and WI-38 VA13 human cell lines |
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Authors: | M T Cushion J J Ruffolo M J Linke P D Walzer |
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Institution: | 2. Department of Anatomy and Cell Biology, College of Medicine, University of Cincinnati, 231 Bethesda Avenue, Cincinnati, Ohio 45267, U.S.A. |
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Abstract: | 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. |
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Keywords: | Protozoa parastic Tissue culture Lung cell lines human Acquired immune deficiency syndrome (AIDS) Dulbecco's Modified Eagle's Medium (DMEM) Fetal bovine serum (FBS) Phosphate-buffered saline (PBS) Eagle's Minimum Essential Medium with Hank's salts (HMEM) |
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