scholarly journals Plasmodium falciparum in vitro continuous culture conditions: A comparison of parasite susceptibility and tolerance to anti-malarial drugs throughout the asexual intra-erythrocytic life cycle

2017 ◽  
Vol 7 (3) ◽  
pp. 295-302 ◽  
Author(s):  
Sandra Duffy ◽  
Vicky M. Avery
Keyword(s):  
Plasmodium Falciparum ◽  
Life Cycle ◽  
Continuous Culture ◽  
Culture Conditions

scholarly journals Interaction of Plasmodium falciparum apicortin with α- and β-tubulin is critical for parasite growth and survival

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Malabika Chakrabarti ◽  
Nishant Joshi ◽  
Geeta Kumari ◽  
Preeti Singh ◽  
Rumaisha Shoaib ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Life Cycle ◽  
Specific Protein ◽  
Parasite Growth ◽  
Growth And Survival ◽  
Apicomplexan Parasites ◽  
Parasite Replication ◽  
Main Components ◽  
Β Tubulin

AbstractCytoskeletal structures of Apicomplexan parasites are important for parasite replication, motility, invasion to the host cell and survival. Apicortin, an Apicomplexan specific protein appears to be a crucial factor in maintaining stability of the parasite cytoskeletal assemblies. However, the function of apicortin, in terms of interaction with microtubules still remains elusive. Herein, we have attempted to elucidate the function of Plasmodium falciparum apicortin by monitoring its interaction with two main components of parasite microtubular structure, α-tubulin-I and β-tubulin through in silico and in vitro studies. Further, a p25 domain binding generic drug Tamoxifen (TMX), was used to disrupt PfApicortin-tubulin interactions which led to the inhibition in growth and progression of blood stage life cycle of P. falciparum.

Continuous Cultures of Plasmodium Falciparum Established in Tanzania From Patients With Acute Malaria

2021 ◽  
Author(s):  
Florence Humphrey Urio ◽  
Matilda Mkombachepa ◽  
Gration Rwegasira ◽  
Twilumba Makene ◽  
Billy Ngasala ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Continuous Culture ◽  
Drug Sensitivity ◽  
Culture Media ◽  
Experimental System ◽  
Dar Es Salaam ◽  
Clinical Samples ◽  
Host Cell Interactions ◽  
Set Up

Abstract BackgroundMalaria morbidity and mortality, almost entirely from Plasmodium falciparum, are still rampant in Africa: therefore, it is important to study the biology of the parasite and the parasite-host cell interactions. In vitro cultivation of Plasmodium falciparum is most useful for this purpose, as well as for investigating drug resistance and possible new therapies. Here we report that the Trager & Jensen continuous culture of P. falciparum can be established in a laboratory in Tanzania with minimal facilities and with modest expenditure.MethodsAn in vitro set-up of continuous culture of P. falciparum was carried out in 2016 to 2020 at Muhimbili university of health and allied sciences, Dar-es salaam. Parasite samples were obtained from patients with acute malaria, frozen parasites and live cultures. Data was collected and analyzed using GraphPad Prism version 8.ResultsWe have successfully achieved exponential growth of existing strains that are used worldwide, as well as of parasites in clinical samples from patients with acute malaria. In the aim to optimize growth we have compared human serum and bovine serum albumin as components of the culture media. In addition, culture synchronization has been achieved using sorbitol.ConclusionThis experimental system is now available to our institution and to researchers aiming at investigating drug sensitivity and mechanisms of protection against Plasmodium falciparum that accrue from various genes expressed in red cells.

scholarly journals In vitro life cycle of pentamidine-resistant amastigotes: stability of the chemoresistant phenotypes is dependent on the level of resistance induced.

1997 ◽  
Vol 41 (9) ◽  
pp. 1898-1903 ◽  
Author(s):  
D Sereno ◽  
J L Lemesre
Keyword(s):  
Life Cycle ◽  
Culture Conditions ◽  
Cross Resistance ◽  
Diminazene Aceturate ◽  
Drug Pressure ◽  
Mouse Macrophages ◽  
Wild Type Clone ◽  
Different Levels

Using a continuous drug pressure protocol, we induced pentamidine resistance in an active and dividing population of amastigote forms of Leishmania mexicana. We selected in vitro two clones with different levels of resistance to pentamidine, with clone LmPENT5 being resistant to 5 microM pentamidine, while clone LmPENT20 was resistant to 20 microM pentamidine. Resistance indexes (50% inhibitory concentration [IC50] after drug presure/IC50 before drug pressure) of 2 (LmPENT5) and 6 (LmPENT20) were determined after drug selection. Both resistant clones expressed significant cross-resistance to diminazene aceturate and primaquine. Pentamidine resistance was not reversed by verapamil, a calcium channel blocker known to reverse multidrug resistance (A. J. Bitonti, et al., Science 242:1301-1303, 1988; A. R. C. Safa et al., J. Biol. Chem. 262:7884-7888, 1987). No difference in the in vitro infectivity for resident mouse macrophages was observed between the wild-type clone (clone LmWT) and pentamidine-resistant clones. During in vitro infectivity experiments, when the life cycle was performed starting from the intramacrophagic amastigote stage, the drug resistance of the resulting LmPENT20 amastigotes was preserved even if the intermediate promastigote stage could not be considered resistant to 20 microM pentamidine. In the same way, when a complete developmental sequence of L. mexicana was achieved axenically by manipulation of appropriate culture conditions, the resulting axenically grown LmPENT20 amastigotes remained pentamidine resistant, whereas LmPENT5 amastigotes lost their ability to resist pentamidine, with IC50s and index of resistance values close to those for the LmWT clone. These results strongly indicate that the level of pentamidine tolerated by resistant amastigotes after the life cycle was dependent on the induced level of resistance. This fact could be significant in the in vivo transmission of drug-resistant parasites by Phlebotominae. Particular attention should be given to the finding that the emergence of parasite resistance is a potential risk of the use of inadequate doses as therapy in humans.

Plasmodium falciparum: Drug sensitivity in vitro of isolates before and after adaptation to continuous culture

1983 ◽  
Vol 56 (1) ◽  
pp. 9-14 ◽  
Author(s):  
J. Le Bras ◽  
P. Deloron ◽  
A. Ricour ◽  
B. Andrieu ◽  
J. Savel ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Continuous Culture ◽  
Drug Sensitivity ◽  
Before And After

scholarly journals Study of some parameters affecting the in vitro cultivation of Plasmodium falciparum within saimiri sciureus red blood cells

1986 ◽  
Vol 81 (2) ◽  
pp. 165-170 ◽  
Author(s):  
T. Fandeur ◽  
J. P. Dedet
Keyword(s):  
Plasmodium Falciparum ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Osmotic Fragility ◽  
Culture Conditions ◽  
Saimiri Sciureus ◽  
In Vitro Cultivation ◽  
In Vitro Growth ◽  
Serum Supplement

The in vitro growth and multiplication of the erythrocytic stages of Plasmodium falciparum within Saimiri sciureus (squirrel monkey) red blood cells have been studied. Various parameters, such as the origin of the red blood cells and serum supplement, nature of the buffer, influence of the final pH of the medium, role of proteose peptone and glucose addition, were investigated. The selection of the best culture conditions led to the obtention of a reproducible in vitro growth of two parasite cycles in Saimiri erythrocytes, which is an useful achievement for in vitro studies. Our failure to establish a continuous culture line for longer than 19 days, could be explained by a dramatic increasing of osmotic fragility of the Saimiri red blood cells related to their small size.

scholarly journals CONTINUOUS CULTURES OF PLASMODIUM FALCIPARUM ESTABLISHED IN TANZANIA FROM PATIENTS WITH ACUTE MALARIA

2021 ◽  
Vol 13 (1) ◽  
Author(s):  
Florence Urio ◽  
Matilda Mkombachepa ◽  
Gration Rwegasira ◽  
Twilumba Makene ◽  
Billy Ngasala ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Continuous Culture ◽  
Drug Sensitivity ◽  
Culture Media ◽  
Experimental System ◽  
Clinical Samples ◽  
In Vitro Cultivation ◽  
Host Cell Interactions ◽  
Set Up

Background: Malaria morbidity and mortality, almost entirely from Plasmodium falciparum, are still rampant in Africa: therefore, it is important to study the biology of the parasite and the parasite-host cell interactions. In vitro cultivation of Plasmodium falciparum is most useful for this purpose, as well as for investigating drug resistance and possible new therapies. Here we report that the Trager & Jensen continuous culture of P. falciparum can be established in a laboratory in Tanzania with minimal facilities and with modest expenditure. Methodology: This was an in-vitro set up of continuous culture of Plasmodium falciparum study, carried out in 2016-2020 at Muhimbili university of health and allied sciences, Dar-es salaam. Parasite samples were obtained from patients with acute malaria, frozen parasites, and live cultures. Data was collected and analyzed using GraphPad Prism version 8. Results: We have successfully achieved exponential growth of existing strains that are used worldwide, as well as of parasites in clinical samples from patients with acute malaria. In the aim to optimize growth we have compared human serum and bovine serum albumin as components of the culture media. Additionally, culture synchronization has been achieved using sorbitol. Conclusion: This experimental system is now available to our institution and to researchers aiming at investigating drug sensitivity and mechanisms of protection against Plasmodium falciparum that accrue from various genes expressed in red cells.  

scholarly journals Insights Gained fromP. falciparumCultivation in Modified Media

2013 ◽  
Vol 2013 ◽  
pp. 1-6 ◽  
Author(s):  
Sanjay A. Desai
Keyword(s):  
Plasmodium Falciparum ◽  
Basic Research ◽  
Culture Conditions ◽  
Parasite Growth ◽  
Human Malaria ◽  
Parasite Culture ◽  
Comparison Of Results ◽  
Modified Media

In vitrocultivation ofPlasmodium falciparum, the agent of severe human malaria, has enabled advances in basic research and accelerated the development of new therapies. Since the introduction ofin vitroparasite culture nearly 40 years ago, most workers have used a medium consisting of RPMI 1640 medium supplemented with lipids and hypoxanthine. While these standardized conditions yield robust parasite growth and facilitate comparison of results from different studies, they may also lead to implicit assumptions that limit future advances. Here, I review recent studies that used modified culture conditions to challenge these assumptions and explore parasite physiology. The findings are relevant to understandingin vivoparasite phenotypes and the prioritization of antimalarial targets.

Plasmodium falciparum: Modifications of the In vitro Culture Conditions Improving Parasitic Yields

1982 ◽  
Vol 68 (6) ◽  
pp. 1072 ◽  
Author(s):  
J. Werner Zolg ◽  
Alexander J. MacLeod ◽  
Ida H. Dickson ◽  
John G. Scaife
Keyword(s):  
Plasmodium Falciparum ◽  
In Vitro Culture ◽  
Culture Conditions

Neutrophil migration through in vitro interstitial matrix

1992 ◽  
Vol 50 (1) ◽  
pp. 894-895
Author(s):  
J. Roemer ◽  
S.R. Simon
Keyword(s):  
Extracellular Matrix ◽  
Smooth Muscle ◽  
Cell Migration ◽  
Smooth Muscle Cells ◽  
Inflammatory Cell ◽  
Neutrophil Migration ◽  
Culture Conditions ◽  
Aortic Smooth Muscle ◽  
Specific Culture

We are developing an in vitro interstitial extracellular matrix (ECM) system for study of inflammatory cell migration. Falcon brand Cyclopore membrane inserts of various pore sizes are used as a support substrate for production of ECM by R22 rat aortic smooth muscle cells. Under specific culture conditions these cells produce a highly insoluble matrix consisting of typical interstitial ECM components, i.e.: types I and III collagen, elastin, proteoglycans and fibronectin.

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