The use of Toxoplasma gondii tachyzoites produced in HeLa cells adhered to Cytodex 1 microcarriers as antigen in serological assays: an application of microcarrier technology
Abstract:Toxoplasma gondii can infect nearly all warm-blooded animals, including humans. In the laboratory diagnosis of toxoplasmosis, serological tests have importance in detecting antibody response. Traditionally T. gondii tachyzoites grown in vivo are being used as an antigen source in serological assays. Currently, tachyzoites produced in vitro are being tested as an antigen source in order to decrease animal use. Microcarrier technology allowed us to grow anchorage-dependent host cells on microcarrier suspension i… Show more
“…In vitro cell culture methods are well established for T . gondii , with various studies published using several distinct cell lines most commonly using tachyzoite stages for infection [ 42 – 44 ]. Alternatively, other T .…”
Section: Review Of Existing In Vitro Models For Protozoan Parasitesmentioning
Protozoan parasites are responsible for severe disease and suffering in humans worldwide. Apart from disease transmission via insect vectors and contaminated soil, food, or water, transmission may occur congenitally or by way of blood transfusion and organ transplantation. Several recent outbreaks associated with fresh produce and potable water emphasize the need for vigilance and monitoring of protozoan parasites that cause severe disease in humans globally. Apart from the tropical parasite Plasmodium spp., other protozoa causing debilitating and fatal diseases such as Trypanosoma spp. and Naegleria fowleri need to be studied in more detail. Climate change and socioeconomic issues such as migration continue to be major drivers for the spread of these neglected tropical diseases beyond endemic zones. Due to the complex life cycles of protozoa involving multiple hosts, vectors, and stringent growth conditions, studying these parasites has been challenging. While in vivo models may provide insights into host–parasite interaction, the ethical aspects of laboratory animal use and the challenge of ready availability of parasite life stages underline the need for in vitro models as valid alternatives for culturing and maintaining protozoan parasites. To our knowledge, this review is the first of its kind to highlight available in vitro models for protozoa causing highly infectious diseases. In recent years, several research efforts using new technologies such as 3D organoid and spheroid systems for protozoan parasites have been introduced that provide valuable tools to advance complex culturing models and offer new opportunities toward the advancement of parasite in vitro studies. In vitro models aid scientists and healthcare providers in gaining insights into parasite infection biology, ultimately enabling the use of novel strategies for preventing and treating these diseases.
“…In vitro cell culture methods are well established for T . gondii , with various studies published using several distinct cell lines most commonly using tachyzoite stages for infection [ 42 – 44 ]. Alternatively, other T .…”
Section: Review Of Existing In Vitro Models For Protozoan Parasitesmentioning
Protozoan parasites are responsible for severe disease and suffering in humans worldwide. Apart from disease transmission via insect vectors and contaminated soil, food, or water, transmission may occur congenitally or by way of blood transfusion and organ transplantation. Several recent outbreaks associated with fresh produce and potable water emphasize the need for vigilance and monitoring of protozoan parasites that cause severe disease in humans globally. Apart from the tropical parasite Plasmodium spp., other protozoa causing debilitating and fatal diseases such as Trypanosoma spp. and Naegleria fowleri need to be studied in more detail. Climate change and socioeconomic issues such as migration continue to be major drivers for the spread of these neglected tropical diseases beyond endemic zones. Due to the complex life cycles of protozoa involving multiple hosts, vectors, and stringent growth conditions, studying these parasites has been challenging. While in vivo models may provide insights into host–parasite interaction, the ethical aspects of laboratory animal use and the challenge of ready availability of parasite life stages underline the need for in vitro models as valid alternatives for culturing and maintaining protozoan parasites. To our knowledge, this review is the first of its kind to highlight available in vitro models for protozoa causing highly infectious diseases. In recent years, several research efforts using new technologies such as 3D organoid and spheroid systems for protozoan parasites have been introduced that provide valuable tools to advance complex culturing models and offer new opportunities toward the advancement of parasite in vitro studies. In vitro models aid scientists and healthcare providers in gaining insights into parasite infection biology, ultimately enabling the use of novel strategies for preventing and treating these diseases.
“…Prior to this, cells derived from other human cells could only survive for a few days in culture, but the behavior of Lacks’s tumor cells was different. This was the first successful attempt to immortalize and maintain human cells in vitro because the ability of Hela cells made them useful in various biological studies [ 19 , 20 , 21 , 22 , 23 ].…”
Cells are very important to researchers due to their use in various biological studies in in vitro and in vivo settings. This importance stems from the short lifespan of most cells under laboratory conditions, which can pose significant challenges, such as the difficulties associated with extraction from the source tissue, ethical concerns about separating cells from human or animal models, limited cell passage ability, and variation in results due to differences in the source of the obtained cells, among other issues. In general, cells in laboratory conditions can divide into a limited number, known as the Hayflick limit, due to telomere erosion at the end of each cellular cycle. Given this problem, researchers require cell lines that do not enter the senescence phase after a limited number of divisions. This can allow for more stable studies over time, prevent the laborious work associated with cell separation and repeated cultivation, and save time and money in research projects. The aim of this review is to summarize the function and effect of immortalization techniques, various methods, their advantages and disadvantages, and ultimately the application of immortalization and cell line production in various research fields.
“…/blast.ncbi.nlm.nih.gov/Blast.cgi) kontrol edilmiştir. T. gondii pozitif DNA örnekleriyle oluşturulmuştur(16). Öncelikle RE bölgesi tasarlanan dış primerler olan F3 ve B3 primerleri kullanılarak PZR ile izole edilmiştir.…”
Toxoplasma gondii insan ve sıcakkanlı hayvanlarda hastalıklara neden olan bir protozoondur. Bu çalışmanın amacı toksoplazmozis tanısı için T. gondii tekrar bölgesine (RE) özgü primerlerin tasarlanması ile geliştirilen floresans esaslı hızlı Döngü Aracılı İzotermal Amplifikasyon (LAMP) testinin analitik hassasiyetini belirlemektir. Gereç ve Yöntem: Polimeraz Zincir Reaksiyonu (PZR) ile elde edilen T. gondii RE, pCR2.1-TOPO vektörüne klonlanmıştır. Spesifik primerler kullanılarak geliştirilen LAMP testinin analitik hassasiyeti pCR2.1-RE vektörünün seri seyreltmeleri ile belirlenmiştir. Bulgular: T. gondii RE genine özgü tasarlanan primerler ile geliştirilen LAMP testinin analitik hassasiyetinin ≤10 plazmit/reaksiyon olduğu tespit edilmiştir. Sonuç: RE bölgesi T. gondii geni içinde 200-300 kere tekrar ettiğinden dolayı geliştirilen testin 0,05 takizoit/reaksiyon saptama limitine sahip olduğu hesaplanmıştır. LAMP testinin kolay, ucuz, hızlı ve saha koşullarına uygun olması ve diğer moleküler testlerden daha hassas olması nedeniyle umut verici olduğu öngörülmektedir.
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