More productive in vitro culture of Cryptosporidium parvum for better study of the intra- and extracellular phases

Cordón, G. Peréz; Marín, Clotilde; Romero, Desirée; Rosales, César; Moreno, Manuel Sánchez; Rosales, M.J.

doi:10.1590/s0074-02762007005000071

Cited by 8 publications

(12 citation statements)

References 27 publications

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“…This hypothesis has in fact been confirmed by the observations of Perez Cordón et al . (2007), in which HCT-8 cell cultures, for which the medium had not been renewed in 7 days, had a large percent of extracellular stages augmented Perez Cordón et al . (2007).…”

Section: Developmental Stages and Cell-free Culture: Different Pointsmentioning

confidence: 99%

“…Several advances and system improvements have also been reported in the last decade (Perez Cordón et al . 2007; Alcantara Warren et al . 2008; Castellanos-Gonzalez et al .…”

Section: Facts and Trends In Advanced Systems Of Cryptosporidium Cultmentioning

confidence: 99%

“…Perez Cordón et al . (2007) used HCT-8 cell cultures, for which the culture was renewed for seven days, infected with C. parvum sporozoites in RPMI-1640 medium with 10% IFBS, CaCl 2 and MgCl 2 1 m m at pH 7·2 increased parasitism by 71% at 48 h vs 14·5%. An increase in the percentage of extracellular stages was clearly documented (25·3%).…”

Section: Facts and Trends In Advanced Systems Of Cryptosporidium Cultmentioning

confidence: 99%

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The truth aboutin vitroculture ofCryptosporidiumspecies

Karanis

2017

Parasitology

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Cryptosporidium research has focused on the development of infection control, and effective therapy that has thus far been hampered by the inability to culture Cryptosporidium in vitro. Other limitations include inadequate animal models, cumbersome screening procedures for chemotherapeutic approaches and a lack of tools for genetic manipulation. These limitations can, however, be eased by the improvement and focused development of in vitro cultivation. The ability to culture relevant Cryptosporidium isolates in vitro and to propagate the life cycle stages that are responsible for causing disease in an infected host is still a critical link. This ability will facilitate other relevant approaches, e.g., the ability to knockout genes and the application of broader screening for drug discoveries and vaccine developments, in combination with new discoveries on the parasite's basic biology, genetic manipulation and new life cycle stages. Success in this effort represents an essential step towards significant progress in the control of cryptosporidiosis.

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Section: Developmental Stages and Cell-free Culture: Different Pointsmentioning

confidence: 99%

“…Several advances and system improvements have also been reported in the last decade (Perez Cordón et al . 2007; Alcantara Warren et al . 2008; Castellanos-Gonzalez et al .…”

Section: Facts and Trends In Advanced Systems Of Cryptosporidium Cultmentioning

confidence: 99%

Section: Facts and Trends In Advanced Systems Of Cryptosporidium Cultmentioning

confidence: 99%

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The truth aboutin vitroculture ofCryptosporidiumspecies

Karanis

2017

Parasitology

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“…The final supplemented medium, consisting of RPMI 1640 with 10% FBS, 15 mM HEPES, 50 mM glucose, and 35 mg of ascorbic acid, 1.0 mg of folic acid, 4.0 mg of 4-aminobenzoic acid, 2.0 mg of calcium pantothenate, 0.1 U of insulin, 100 U of penicillin G, 100 mg of streptomycin, and 0.25 mg of amphotericin B per ml (pH 7.4) supported a ten-fold increase in parasite abundance compared to base RPMI-1640 (Upton et al, 1995). This medium is commonly employed when culturing C. parvum, although another medium formulation has been developed which can maintain HCT-8 cells and parasitism for two weeks (Perez Cordon et al, 2007). This formulation consists of RPMI-1640 medium with 10% FBS, pH 7.2 with CaCl 2 and MgCl 2 at 1 mM, and also requires that the cells (HCT-8) have no medium renewal for a week prior to infection.…”

Section: Cryptosporidium Culturingmentioning

confidence: 99%

Past and future trends of Cryptosporidium in vitro research

Bones

Jossé

et al. 2019

Experimental Parasitology

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Cryptosporidium is a genus of single celled parasites capable of infecting a wide range of animals including humans. Cryptosporidium species are members of the phylum apicomplexa, which includes well-known genera such as Plasmodium and Toxoplasma. Cryptosporidium parasites cause a severe gastro-intestinal disease known as cryptosporidiosis. They are one of the most common causes of childhood diarrhoea worldwide, and infection can have prolonged detrimental effects on the development of children, but also can be life threatening to HIV/AIDS patients and transplant recipients. A variety of hosts can act as reservoirs, and Cryptosporidium can persist in the environment for prolonged times as oocysts. While there has been substantial interest in these parasites, there is very little progress in terms of treatment development and understanding the majority of the life cycle of this unusual organism. In this review, we will provide an overview on the existing knowledge of the biology of the parasite and the current progress in developing in vitro cultivation systems. We will then describe a synopsis of current and next generation approaches that could spearhead further research in combating the parasite.

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“…Human colorectal carcinoma (HCT-8) and Madin Darby Canine Kidney (MDCK) cells have both shown promise during the previous decades of C. parvum experiments, yet often fall short due either to rapid culture senescence or an inability to properly cultivate the range of endogenous phases of C. parvum noted in the medical literature (Arrowood, 2002;Girouard, Gallant, Akiyoshi, Nunnari, & Tzipori, 2006;Hijjawi, Meloni, Morgan, & Thompson, 2001;Upton, Tilley, & Brillhart, 1994). Besides the choice of cell subtype, efforts to improve media formulation have led to prolonged culture time of the host, allowing the parasite to differentiate into further developmental stages or even allowing multiple rounds of infection (Miller et al, 2018a;Morada et al, 2016;Perez Cordon et al, 2007). Prior to developing this system, several cancerous lines-isolated from a range of cancerous tissues-were tested for their propensity to infection (see Miller et al, 2018a).…”

Section: Background Informationmentioning

confidence: 99%

A Cell Culture Platform for the Cultivation of Cryptosporidium parvum

et al. 2019

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Cryptosporidium is a genus of ubiquitous unicellular parasites belonging to the phylum Apicomplexa. Cryptosporidium species are the second largest cause of childhood diarrhea and are associated with increased morbidity. Accompanying this is the low availability of treatment and lack of vaccines. The major barrier to developing effective treatment is the lack of reliable in vitro culture methods. Recently, our lab has successfully cultivated C. parvum in the esophageal cancer–derived cell line COLO‐680N, and has been able to maintain infection for several weeks. The success of this cell line was assessed with a combination of various techniques including fluorescent microscopy and qPCR. In addition, to tackle the issue of long‐term oocyst production in vitro, a simple, low‐cost bioreactor system using the COLO‐680N cell line was established, which produced infectious oocysts for 4 months. This chapter provides details on the methodologies used to culture, maintain, and assess Cryptosporidium infection and propagation in COLO‐680N. © 2019 by John Wiley & Sons, Inc.

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More productive in vitro culture of Cryptosporidium parvum for better study of the intra- and extracellular phases

Cited by 8 publications

References 27 publications

The truth aboutin vitroculture ofCryptosporidiumspecies

The truth aboutin vitroculture ofCryptosporidiumspecies

Past and future trends of Cryptosporidium in vitro research

A Cell Culture Platform for the Cultivation of Cryptosporidium parvum

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