Toxoplasma tachyzoites from cell culture are more appropriate in some situations

2010 ◽  
Vol 63 (5) ◽  
pp. 438-440
Author(s):  
Jean M W Chatterton ◽  
Susan McDonagh ◽  
Darrel O Ho-Yen

BackgroundLaboratories traditionally culture toxoplasma tachyzoites in animals for testing and experimental use. This article considers why available cell culture methods are not used more often.AimTo compare HeLa cell culture and animal culture for production of toxoplasma tachyzoites.MethodsIn 2000 HeLa culture replaced animal culture for continuous production of toxoplasma tachyzoites in the Scottish Toxoplasma Reference Laboratory. The performance of animal culture (1994–1998) was compared with HeLa culture (2004–2008). A PubMed search was carried out for 1998 and 2008 to assess the culture methods used in laboratories.ResultsAnimal culture was able to produce higher yields of tachyzoites (109 from a cotton rat peritoneal harvest compared to 107 from a 75 cm2 cell culture flask) but significantly more HeLa cultures were successful (93% versus 84%; p=0.025). There was no difference in the quality of tachyzoites from animal and HeLa cultures as demonstrated by the high levels of success in the dye test. HeLa culture offered significant advantages in flexibility and control. A review of the literature showed no significant change in the method of culture used in laboratories between 1998 and 2008 (p=0.36).ConclusionThe availability of cell culture methods and the increasingly stringent regulations on the use of animals have not resulted in a decline in the use of animal culture. Animals are necessary for certain experiments but many studies could use cell-culture-derived parasites.

1992 ◽  
Vol 20 (1) ◽  
pp. 52-60
Author(s):  
Gabriela Ciapetti ◽  
Elisabetta Cenni ◽  
Daniela Cavedagna ◽  
Loredana Pratelli ◽  
Arturo Pizzoferrato

Cell culture techniques are usually used in the field of biomaterials research and development in order to detect toxic components. Morphological assays are the most widely used methods and give the very first information about the biological compatibility of materials. Cell function assays give more quantitative data, but the comparison of data between different laboratories is difficult. Some of the cell culture methods that are used for biocompatibility studies are described briefly here, and results from our laboratory are reported. Despite some inherent limitations of the cell culture techniques, they are an accurate and reliable method of predicting the biological compatibility of materials to be implanted in vivo.


2021 ◽  
Vol 129 (Suppl_1) ◽  
Author(s):  
Selen Uman ◽  
Jason A Burdick

Introduction: Early studies have shown therapeutic benefits of mesenchymal stromal cells (MSCs) in cardioprotection due to their angiogenic, proliferative, anti-apoptotic and anti-inflammatory properties, which are now attributed to secreted factors such as extracellular vesicles (EVs). While MSC-EVs have shown promise in small animals for cardiovascular therapies, large animal studies are required to evaluate the therapeutic benefit of MSC-EVs for clinical translation. One of the biggest challenges for large animal studies is the need to generate clinically-relevant quality and quantity of EVs without batch-to-batch variations that could compromise efficacy. This study aims to explore three different cell culture methods (traditionally-used tissue culture plates (TCP), 3-D printed bioscaffolds in a perfusion system (P), and microcarriers in dynamic spinner flask conditions (M)) to scale-up the production of MSC-EVs across four different biological donors and rigorously investigate EV yield, size, shape, and content. Methods: MSCs were isolated from the iliac crest of four different Yucatan minipigs using heparinized syringes, and cells were expanded to passage four, at which point they were seeded onto the respective cell culture methods. EVs were collected from conditioned medium (CM) via differential ultracentrifugation. EV size, distribution, yield, and protein concentration were studied using Nanoparticle Tracking Analysis (NTA) and microBCA assays. Results: Both perfusion bioreactor and spinner flask systems enabled sustained maintenance of large numbers of cells. Across biological donors and fabrication methods, modes remained within 50-150 nm and were not statistically different. Microcarrier-based spinner flasks and perfusion bioreactor set-ups both improved EV yield, up to 6 times in efficiency. Ongoing research focuses on examining differences in EV content across biological donors using RNA-sequencing and proteomics.


2018 ◽  
Vol 42 (2) ◽  
pp. 257-267
Author(s):  
Jean-René Arseneau ◽  
Chantal Gautreau ◽  
Linda Boston ◽  
Michel L. Goguen ◽  
Mark Laflamme

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