Plasmodium falciparum: One-step growth in a semi-defined medium and the stimulatory effect of human seric lipoproteins and liposomes

Nivet, C; Guillotte, Micheline; Silva, Luiz Pereira da

doi:10.1016/0014-4894(83)90008-5

Cited by 11 publications

(5 citation statements)

References 13 publications

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“…Continuous in vitro culture of P. falciparum was obtained in medium containing unsaturated fatty acids adsorbed on serum albumin but the parasitemia was very low in comparison to medium with human serum (Willet and Canfield, 1984). One cycle of growth was obtained in vitro using a medium supplemented with human lipoprotein fractions and dialysable serum factors (Nivet et al, 1983).…”

mentioning

confidence: 99%

Lipid traffic between high density lipoproteins and Plasmodium falciparum-infected red blood cells.

Grellier

Rigomier

Clavey

et al. 1991

The Journal of Cell Biology

133

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Abstract. Several intraerythrocytic growth cycles of Plasmodium falciparum could be achieved in vitro using a serum free medium supplemented only with a human high density lipoprotein (HDL) fraction (d = 1.063-1.210). The parasitemia obtained was similar to that in standard culture medium containing human serum. The parasite development was incomplete with the low density lipoprotein (LDL) fraction and did not occur with the VLDL fraction. The lipid traffic from HDL to the infected erythrocytes was demonstrated by pulse labeling experiments using HDL loaded with either fluorescent NBD-phosphatidylcholine (NBD-PC) or radioactive [aH]palmitoyl-PC. At 37°C, the lipid probes rapidly accumulated in the infected cells. After incubation in HDL medium containing labeled PC, a subsequent incubation in medium with either an excess of native HDL or 20% human serum induced the disappearance of the label from the erythrocyte plasma membrane but not from the intraerythrocytic parasite. Internalization of lipids did not occur at 4°C. The mechanism involved a unidirectional flux of lipids but no endocytosis. The absence of labeling of P. falciparum, with HDL previously [125I]iodinated on their apolipoproteins or with antibodies against the apolipoproteins AI and All by immunofluorescence and immunoblotting, confirmed that no endocytosis of the HDL was involved. A possible pathway of lipid transport could be a membrane flux since fluorescence videomicroscopy showed numerous organdies labeled with NBD-PC moving between the erythrocyte and the parasitophorous membranes. TLC analysis showed that a partial conversion of the PC to phosphatidylethanolamine was observed in P. falciparum-infected red cells after pulse with [3H]palmitoyl-PC-HDL. The intensity of the lipid traffic was stage dependent with a maximum at the trophozoite and young schizont stages (38 t~ h of the erythrocyte life cycle). We conclude that the HDL fraction appears to be a major lipid source for Plasmodium growth.URING its intraerythrocytic phase, the human malarial parasite Plasmodium falciparum reproduces at a rapid rate, completing its development from the infective merozoite to the mature schizont (16-20 nuclei) in <48 h. After the invasion step, the parasite is isolated from the erythroeyte cytoplasm by a parasitophorous vacuole membrane and undergoes a sequential development through a ring form, trophozoite, schizont, and finally the differentiation of 10-20 merozoites. The new merozoites are released into the bloodstream by erythrocyte bursting.Throughout the schizogonic phase, the parasite sequential development that takes place, corresponds to considerable nuclear and cytoplasmic transformations, including a fivefold increase of total phospholipid content and a decrease in the cholesterol-phospholipid ratio of the infected cell (Holz, 1977;Sherman, 1979;Vial et al., 1982a). Studies of the fatty acid composition have shown that the ability of P/asmodium to perform saturation or desaturation reactions of aliphatic chains, as well as chain lengthening and s...

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mentioning

confidence: 99%

Lipid traffic between high density lipoproteins and Plasmodium falciparum-infected red blood cells.

Grellier

Rigomier

Clavey

et al. 1991

The Journal of Cell Biology

133

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“…There are obvious advantages of using serum replacements: limited batch-tobatch variation, which is a prerequisite for a standardized protocol; compatibility with any human blood type; production at industrial scale and availability through commercial sources, usually at lower costs than non-immune human serum; ease in handling and transport; limited risks of blood-borne pathogens, such as human immunodeficiency virus and hepatitis virus. Serum replacements with which in vitro cultivation of P. falciparum has been achieved are listed in Table 3 [54,99,101,122,155,156,167,174,[185][186][187][188][189][190][191][192][193][194][195][196][197][198][199]. Good growth at 48 h with 0.5 mg/mL HDL or 1.8 mg/mL LDL; long-term culture not performed [187] 0.25-0.5 mg/mL HDL fraction Successful short-term culture comparable to 10% human serum [189] GF21 Ammonium sulfate fraction of adult bovine serum, insulin, transferrin, ethanolamine, sodium selenite Very good growth in the GIT medium 4 ; successful long-term culture [101] Nutridoma-SR ® 1 Albumin, insulin, transferrin, cholesterol, organic and inorganic compounds At 4% (v/v), growth comparable with serum-supplemented RPMI; may be strain-dependent [190] Nutridoma alone at 4%, poor growth; 1% Nutridoma + 0.25% or 0.5% Albumax ® , very good growth [191] Ultroser ® G 2 Binding proteins, fatty acids, phospholipids, adhesion factors, hormones, vitamins, growth factors 1-4% Ultroser, no growth; ≤0.5% Ultroser + ≤2% human serum, moderate growth for 4 days [192] 2-4% Ultroser, no growth; 0.5-1% low growth on days 5-9 [193] Albumax I or II 3 Lipid-enriched bovine serum albumin…”

Section: Serum-free Culturementioning

confidence: 99%

“…There are obvious advantages of using serum replacements: limited batch-to-batch variation, which is a prerequisite for a standardized protocol; compatibility with any human blood type; production at industrial scale and availability through commercial sources, usually at lower costs than non-immune human serum; ease in handling and transport; limited risks of blood-borne pathogens, such as human immunodeficiency virus and hepatitis virus. Serum replacements with which in vitro cultivation of P. falciparum has been achieved are listed in Table 3 [ 54 , 99 , 101 , 122 , 155 , 156 , 167 , 174 , 185 , 186 , 187 , 188 , 189 , 190 , 191 , 192 , 193 , 194 , 195 , 196 , 197 , 198 , 199 ].…”

Section: Serum and Serum Substitutesmentioning

confidence: 99%

“…One of the key steps necessary to attain the serum-free culture of P. falciparum is to identify human serum component(s) required to satisfy the parasites’ nutritional needs. In short-term experiments with a laboratory-adapted strain grown in a “basic” RPMI 1640 medium (supplemented with fatty acid-free bovine serum albumin (5 mg/mL) and 10% dialyzable human serum factors), parasites did not grow even for 48 h in the absence of nondialyzable macromolecules [ 187 ]. The addition of seric lipoproteins (low-density lipoproteins (LDLs) and high-density lipoproteins (HDLs)) to the medium supported parasite growth, but not to the extent of the control medium with 10% human serum.…”

Section: Serum and Serum Substitutesmentioning

confidence: 99%

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Cultivation of Asexual Intraerythrocytic Stages of Plasmodium falciparum

Basco

2023

Pathogens

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Successfully developed in 1976, the continuous in vitro culture of Plasmodium falciparum has many applications in the field of malaria research. It has become an important experimental model that directly uses a human pathogen responsible for a high prevalence of morbidity and mortality in many parts of the world and is a major source of biological material for immunological, biochemical, molecular, and pharmacological studies. Until present, the basic techniques described by Trager and Jensen and Haynes et al. remain unchanged in many malaria research laboratories. Nonetheless, different factors, including culture media, buffers, serum substitutes and supplements, sources of erythrocytes, and conditions of incubation (especially oxygen concentration), have been modified by different investigators to adapt the original technique in their laboratories or enhance the in vitro growth of the parasites. The possible effects and benefits of these modifications for the continuous cultivation of asexual intraerythrocytic stages of P. falciparum, as well as future challenges in developing a serum-free cultivation system and axenic cultures, are discussed.

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“…Although numerous studies have attempted to identify the factors and substances able to sustain parasite growth (Asahi & Kanazawa, 1994;Asahi et al, 1996Asahi et al, , 2005Cranmer et al, 1997;Divo and Jensen. 1982;Lingnau et al, 1994;Mi-Ichi et al, 2006;Nivet et al, 1983;Ofulla et al, 1993;Willet and Canfield, 1984), the establishment of a fully-defined culture medium for the parasite has represented a major challenge. We previously reported that GFS supported intraerythrocytic growth of P. falciparum (Asahi and Kanazawa, 1994;Asahi et.…”

Section: Cdm For Continuous Intraerythrocytic Growth Of P Falciparummentioning

confidence: 99%

Intraerythrocytic Plasmodium falciparum Growth in Serum-Free Medium with an Emphasis on Growth-Promoting Factors

Asahi¹

2012

Malaria Parasites

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Plasmodium falciparum: One-step growth in a semi-defined medium and the stimulatory effect of human seric lipoproteins and liposomes

Cited by 11 publications

References 13 publications

Lipid traffic between high density lipoproteins and Plasmodium falciparum-infected red blood cells.

Lipid traffic between high density lipoproteins and Plasmodium falciparum-infected red blood cells.

Cultivation of Asexual Intraerythrocytic Stages of Plasmodium falciparum

Intraerythrocytic Plasmodium falciparum Growth in Serum-Free Medium with an Emphasis on Growth-Promoting Factors

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