Morphological comparison of axenic amastigogenesis of trypomastigotes and metacyclic forms of Trypanosoma cruzi

Improved Method for In Vitro Secondary Amastigogenesis ofTrypanosoma cruzi: Morphometrical and Molecular Analysis of Intermediate Developmental Forms

“…The parasites showed dramatic cell restructuring with a gradual and progressive reduction of their size and the readsorption of their flagellum and undulating membrane until they reached the typical round shape of tissue-derived amastigotes (Figure 2). These results are consistent with previous reports that document the existence of several intermediate forms during the life cycle of T. cruzi , either under in vivo or in vitro conditions that showed the same gradual and progressive parasite morphometrical transformation, apparently as an obligatory step for the differentiation process [1, 3, 5, 8, 18–22]. The quantitative analysis demonstrated that parasite transformation was initiated immediately after the exposure to hgDMEM at pH5 and 37°C, thus reaching 100% IFs after only 2 hours of incubation (Figure 4); this indicates a faster and more efficient differentiation rate than previously reported [5, 8].…”

“…These results are consistent with previous reports that document the existence of several intermediate forms during the life cycle of T. cruzi , either under in vivo or in vitro conditions that showed the same gradual and progressive parasite morphometrical transformation, apparently as an obligatory step for the differentiation process [1, 3, 5, 8, 18–22]. The quantitative analysis demonstrated that parasite transformation was initiated immediately after the exposure to hgDMEM at pH5 and 37°C, thus reaching 100% IFs after only 2 hours of incubation (Figure 4); this indicates a faster and more efficient differentiation rate than previously reported [5, 8]. These differences could be the result of the absence of BSA in the differentiation medium used in this study in comparison with the 0.4% BSA contained in the previously reported medium [5, 8].…”

“…Several researchers have studied some of the factors that represent physiological stress for the parasite and have demonstrated that temperature, nutritional conditions, and pH stimulate morphological differentiation during amastigogenesis [5–8]. …”

“…The vast majority of the information regarding in vivo and in vitro amastigogenesis comes from data in which tissue-derived trypomastigotes were used because it was possible to obtain higher yields of these parasites [3, 5, 8–15]. Ultra-structural and molecular analyses during the trypomastigote to amastigote transformation have shown a complex and progressive morphological rearrangement of parasite shape and flagellum that has been associated with the differential expression of stage-specific antigens [3, 5, 8].…”

Detection of Pathogens Using Microfluidics and Biosensors

“…For instance, RBCs have a diameter between 6.2 and 8.2 μm and a thickness between 0.8 and 1 μm [38], while the parasite has a length between 15 and 24 μm [39] and a width of approximately 1.09 μm [40]. These differences in size and density suggest the possibility of using a microfluidic device as a separation platform.…”