Long-term in vitro cultures of Plasmodium berghei and preliminary observations on gametocytogenesis

Janse, Chris J.; Mons, Barend; Croon, J.J.A.B.; Káay, H. J. Van Der

doi:10.1016/0020-7519(84)90083-3

Cited by 48 publications

(15 citation statements)

References 10 publications

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“…the number of nuclei in schizonts, was observed after three life cycles of the parasite. This suggested insufficiency in nutrient levels, which was not overcome by increasing FCS concentration to more than 5% [13]. Medium composition was modified by addition of DMEM/F12 to RPMI1640.…”

Section: Resultsmentioning

confidence: 99%

“…The pellet was diluted at 2.5% haematocrit with fresh medium. Non-infected red blood cells (NRBC) were obtained from healthy mice, and were added twice a week after platelets had been carefully removed from the blood by two washes and centrifugation at 400 g. Buffy coat was removed following Janse et al [13]. Culture was conducted in 75 cm 2 flasks, flushed with gas (5% CO 2 , 5% O 2 , 90% N 2 ) and totally sealed.…”

Section: Methodsmentioning

confidence: 99%

“…Starting culture conditions followed those described by Mons et al and Janse et al [6,13]. To avoid mild haemolysis of IRBC in culture, tests were conducted to adapt the density of the medium and the method of shaking.…”

Section: Methodsmentioning

confidence: 99%

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In vitro culture of Plasmodium berghei-ANKA maintains infectivity of mouse erythrocytes inducing cerebral malaria

Jambou

El‐Assaad

Combes

et al. 2011

Malar J

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BackgroundInfection with Plasmodium berghei is a widely used model of murine malaria and a powerful tool for reverse genetic and pathogenesis studies. However, the efficacy of in vitro reinvasion of erythrocytes is generally low, limiting in vitro studies.MethodsPlasmodium berghei ANKA-infected blood obtained from a susceptible infected mouse was cultured in various conditions and in vitro parasitaemia was measured every day to evaluate the rate of reinvasion.ResultsHigh quality culture media were used and reinvasion rates were improved by vigorous orbital shaking of the flask and increasing density of the medium with gelatin.DiscussionUsing these settings, reinvasion of normal mouse erythrocytes by the parasite was obtained in vitro over two weeks with preservation of the infectivity in vivo.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

In vitro culture of Plasmodium berghei-ANKA maintains infectivity of mouse erythrocytes inducing cerebral malaria

Jambou

El‐Assaad

Combes

et al. 2011

Malar J

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“…Gametocytes were counted according to the reported criteria (Janse et al 1984). The number of red blood cells (RBC) was counted using a Thoma hemocytometer.…”

Section: Count For Blood-stage Parasitesmentioning

confidence: 99%

Superoxide-dependent and -independent pathways are involved in the transmission blocking of malaria

Harada

Owhashi

Suguri

et al. 2001

Parasitology Research

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Superoxide plays a crucial role in innate immunity to various pathogens. We examined the role of superoxides in the transmission of malaria using gp91phox knockout (X-CGD) mice that lack the ability to produce superoxide. Mosquitoes that fed on X-CGD mice infected intraperitoneally with Plasmodium berghei NK65 ANKA formed more oocysts than did those that fed on control mice at any day after infection. The number of oocysts peaked on day 5 post-infection in X-CGD and control mice and then decreased significantly after day 5 post-infection. However, on day 7 post-infection, the infectivity of gametocytes in X-CGD mice was significantly higher than that in control mice. These results show that two pathways, superoxide-dependent and -independent, are involved in the host systems regulating the transmission of malaria and inhibiting gametocyte development.

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“…chronized as has been described (11)(12)(13). Parasites synchronized in this way typically have a window of development no greater than 3 h. Blood containing ring forms was collected from the rats, and leukocytes were removed as described and incubated under standard in vitro culture conditions (11) for 21-24 h to allow the development of the rings into mature schizonts.…”

Section: Parasites-mentioning

confidence: 99%

Species-specific Regulation and Switching of Transcription between Stage-specific Ribosomal RNA Genes in Plasmodium berghei

Waters

Spaendonk

Ramesar

et al. 1997

Journal of Biological Chemistry

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Malaria parasites (Plasmodium spp.) differentially express structurally distinct sets of rRNA genes in a stagespecific manner. The four rRNA genes of the rodent malaria parasite, P. berghei, form two classes of 2 units that are genetically unlinked and termed A-type and S-type. Through Northern analysis and in situ hybridization, expression of the units was demonstrated in synchronized parasite preparations covering the developmental pathway from the initiation of the blood-stage asexual cycle to the production of mature ookinetes. A-type units were transcribed in direct response to cell growth in bloodstage asexual parasites yet were differentially regulated during male (inactive) and female (active) gametocytogenesis. S-type expression was not confined solely to the mosquito stages and exhibited a finite period of expression in a subset of bloodstage trophozoites that was significantly elevated in gametocyte-producing parasites. Unlike in the human parasite, P. falciparum, there was no evidence for accumulation of precursor forms of the S-type transcripts in gametocytes. No significant rRNA transcription was observed in cultured, fertilized ookinetes until ϳ20 h of development when S-type transcription was initiated. The results further demonstrate that in Plasmodium the expression of the different rRNA units is linked to developmental progression but in a species-specific manner.

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Long-term in vitro cultures of Plasmodium berghei and preliminary observations on gametocytogenesis

Cited by 48 publications

References 10 publications

In vitro culture of Plasmodium berghei-ANKA maintains infectivity of mouse erythrocytes inducing cerebral malaria

In vitro culture of Plasmodium berghei-ANKA maintains infectivity of mouse erythrocytes inducing cerebral malaria

Superoxide-dependent and -independent pathways are involved in the transmission blocking of malaria

Species-specific Regulation and Switching of Transcription between Stage-specific Ribosomal RNA Genes in Plasmodium berghei

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