Babesia bigemina : Advances in continuous in vitro culture using serum-free medium supplemented with insulin, transferrin, selenite, and putrescine

Rojas-Martínez, Carmen; Rodríguez-Vivas, Róger Iván; Millán, Julio Vicente Figueroa; Acosta-Viana, Karla Y.; Ruiz, Edwin José Gutiérrez; Garfias, Carlos Ramón Bautista; Lira-Amaya, José J.; Polanco-Martínez, Diego J.; Martínez, Jesús Antonio Álvarez

doi:10.1016/j.parint.2017.11.003

Cited by 22 publications

(33 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Further disadvantages of serum supplementation include batch‐to‐batch variability, high costs and ethical concerns (van der Valk et al., ), making the omission of FBS or newborn calf serum in culture media desirable. Serum‐free cultivation techniques have been reported for related apicomplexan parasites such as Theileria equi , Babesia caballi , Babesia bovis and Babesia bigemina (Rojas Martinez et al., ; Rojas‐Martinez et al., ; Zweygarth, Just, van Niekerk, & de Waal, ; Zweygarth, van Niekerk, & de Waal, ).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Serum‐free in vitro cultivation of Theileria annulata and Theileria parva schizont‐infected lymphocytes

Zweygarth

Nijhof

Knorr

et al. 2020

Transbound Emerg Dis

View full text Add to dashboard Cite

Theileriosis is a tick‐borne disease caused by intracellular protozoa of the genus Theileria. The most important species in cattle are Theileria annulata and Theileria parva. Both species transform leucocyte host cells, resulting in their uncontrolled proliferation and immortalization. Vaccination with attenuated T. annulata‐infected cell lines is currently the only practical means of inducing immunity in cattle. Culture media for Theileria spp. typically contain 10%–20% foetal bovine serum (FBS). The use of FBS is associated with several disadvantages, such as batch‐to‐batch variation, safety and ethical concerns. In this study, the suitability of serum‐free media for the cultivation of Theileria‐transformed cell lines was examined. Three commercial serum‐free media (HL‐1, ISF‐1 and Hybridomed DIF 1000) were evaluated for their ability to support growth of the T. annulata A288 cell line. The generation doubling times were recorded for each medium and compared with those obtained with conventional FBS‐containing RPMI‐1640 medium. ISF‐1 gave the shortest generation doubling time, averaging 35.4 ± 2.8 hr, significantly shorter than the 52.2 ± 14.9 hr recorded for the conventional medium (p = .0011). ISF‐1 was subsequently tested with additional T. annulata strains. The doubling time of a Moroccan strain was significantly increased (65.4 ± 15.9 hr) compared with the control (47.7 ± 7.5 hr, p = .0004), whereas an Egyptian strain grew significantly faster in ISF‐1 medium (43.4 ± 6.5 hr vs. 89.3 ± 24.8 hr, p = .0001). The latter strain also showed an improved generation doubling time of 73.7 ± 21.9 hr in an animal origin‐free, serum‐free, protein‐free medium (PFHM II) compared with the control. Out of four South African T. parva strains and a Theileria strain isolated from roan antelope (Hippotragus equinus), only one T. parva strain could be propagated in ISF‐1 medium. The use of serum‐free medium may thus be suitable for some Theileria cell cultures and needs to be evaluated on a case‐by‐case basis. The relevance of Theileria cultivation in serum‐free media for applications such as vaccine development requires further examination.

show abstract

Section: Introductionmentioning

confidence: 99%

“…Culture media for Theileria spp. et al, 2016;Rojas-Martinez et al, 2018;Zweygarth, Just, van Niekerk, & de Waal, 1997;Zweygarth, van Niekerk, & de Waal, 1999).…”

unclassified

Serum‐free in vitro cultivation of Theileria annulata and Theileria parva schizont‐infected lymphocytes

Zweygarth

Nijhof

Knorr

et al. 2020

Transbound Emerg Dis

View full text Add to dashboard Cite

show abstract

“…When the combination of medium A-DMEM/F12 with ins-trans-sel (2000, 1100, 1.34 mg/L) and Putrescine (Pu, 0.1012 mg/L) was used, maximal PPE of 17.26 and 14.8% were obtained for B. bovis and B. bigemina , respectively. With this culture medium combination, higher growth rates of parasite proliferation were shown ( 92 , 93 ).…”

Section: Elimination Of Bovine Serum In the Culture Medium And Supplementioning

confidence: 93%

“…In Mexico, it was shown that B. bovis and B. bigemina can be adequately controlled for in vitro proliferation in a bovine serum-free medium with supplements such as insuline (ins), transferrine (trans) and selenite (sel) at optimal concentrations ( 91 – 93 ). The Advanced DMEM/F12 culture medium (A-DMEM/F12) supplemented with 40% adult bovine serum (v/v), was selected as an alternate medium to the traditional M-199 with Earle's sales (M-199).…”

Section: Elimination Of Bovine Serum In the Culture Medium And Supplementioning

confidence: 99%

See 1 more Smart Citation

An Overview of Current Knowledge on in vitro Babesia Cultivation for Production of Live Attenuated Vaccines for Bovine Babesiosis in Mexico

2020

View full text Add to dashboard Cite

The instrumentation of the in vitro culture system has allowed researchers to learn more about the metabolic and growth behavior of Babesia spp. The various applications for in vitro cultivation of Babesia include obtaining attenuated strains for vaccination or pre-munition, the selection of pure lines with different degrees of virulence, studies on biological cloning, ultrastructure, antigen production for diagnostics, drug sensitivity assessments, and different aspects of parasite biology. Although there are different types of vaccines that have been tested against bovine babesiosis, so far, the only procedure that has offered favorable results in terms of protection and safety has been the use of live attenuated vaccines. In countries, such as Australia, Argentina, Brazil, Uruguay and Israel, this type of vaccine has been produced and used. The alternative to live vaccines other than splenectomized calf-derived biological material, has been the in vitro cultivation of Babesia bovis and B. bigemina . The development of in vitro culture of Babesia spp. strains in a defined medium has been the basis for the initiation of a source of parasites and exoantigens for a variety of studies on the biochemistry and immunology of babesiosis. The use of live immunogens from attenuated strains derived from in vitro culture is highlighted, which has been proposed as an alternative to control bovine babesiosis. In several studies performed in Mexico, this type of immunogen applied to susceptible cattle has shown the induction of protection against the experimental heterologous strain challenge with both, Babesia -infected blood and animal exposure to confrontations on tick vector-infested farms. The combination of transfection technologies and the in vitro culture system as integrated methodologies would eventually give rise to the generation of genetically modified live vaccines. However, a greater challenge faced now by researchers is the large-scale cultivation of Babesia parasites for mass production and vaccine distribution.

show abstract

Ethyl‐substituted Thioflavin T as a Fluorescent Probe for Detecting the Conformation of Transferrin

Shi

Zhao

et al. 2019

ChemistrySelect

View full text Add to dashboard Cite

Human serum transferrin (hTf) is found that can be used as novel drug active targeted vector, which have broad application prospects in the field of tumor-targeted therapeutics. A new method to recognize human transferrin (hTf) conformations was developed using ThTÀ E. ThTÀ E with three ethyl functional groups has better hydrophobicity than ThT, which can better recognize transferrin and achieve detection of the open conformation of Tf (apo-Tf) against the closed one (holo-Tf). In this study, ThTÀ E and ThT with similar main core structures were selected to evaluate the effect of heterocycles on hTf binding by spectroscopic methods, and a more excellent fluorescent probe ThTÀ E was selected for the recognition of transferrin conformation. And ThTÀ E has a detection limit of 0.27 μM for apo-Tf. This study provides a theoretical basis for ThTÀ E as a biomolecular probe for detecting the mutual conversion of two conformational proteins.

show abstract

Babesia bigemina : Advances in continuous in vitro culture using serum-free medium supplemented with insulin, transferrin, selenite, and putrescine

Cited by 22 publications

References 36 publications

Serum‐free in vitro cultivation of Theileria annulata and Theileria parva schizont‐infected lymphocytes

Serum‐free in vitro cultivation of Theileria annulata and Theileria parva schizont‐infected lymphocytes

An Overview of Current Knowledge on in vitro Babesia Cultivation for Production of Live Attenuated Vaccines for Bovine Babesiosis in Mexico

Ethyl‐substituted Thioflavin T as a Fluorescent Probe for Detecting the Conformation of Transferrin

Contact Info

Product

Resources

About