Simple supplementation of serum-free medium produces gametocytes of Plasmodium falciparum that transmit to mosquitoes
Abstract Background Human serum is a major component of Plasmodium falciparum culture medium, and can be replaced with AlbuMAX™ II, a lipid-rich bovine serum albumin, for asexual cultures. However, gametocytes produced without serum are poorly infective to mosquitoes. Serum suffers from high cost, l...
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| Language: | English |
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BMC
2024-09-01
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| Series: | Malaria Journal |
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| Online Access: | https://doi.org/10.1186/s12936-024-05094-8 |
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| author | Sabyasachi Pradhan Prince Chigozirim Ubiaru Lisa Ranford-Cartwright |
| author_facet | Sabyasachi Pradhan Prince Chigozirim Ubiaru Lisa Ranford-Cartwright |
| author_sort | Sabyasachi Pradhan |
| collection | DOAJ |
| description | Abstract Background Human serum is a major component of Plasmodium falciparum culture medium, and can be replaced with AlbuMAX™ II, a lipid-rich bovine serum albumin, for asexual cultures. However, gametocytes produced without serum are poorly infective to mosquitoes. Serum suffers from high cost, limited availability, and variability in quality. Methods Several commercially-available media supplements were tested for their ability to support parasite growth and production of P. falciparum (3D7) gametocytes in standard RPMI1640 medium containing 0.5% AlbuMAX. The impact on asexual growth and gametocyte production with each supplement was assessed and compared to standard RPMI1640 medium containing 10% human serum, as well as to medium containing 0.5% AlbuMAX alone. The infectivity of gametocytes produced with one supplement to Anopheles gambiae sensu stricto was assessed by standard membrane feeding assay and measuring both prevalence of infection and oocyst intensity. Results Supplementation of medium containing 0.5% AlbuMAX with five supplements did not affect asexual growth of P. falciparum, and four of the five supplements supported early gametocyte production. The supplement producing the highest number of gametocytes, ITS-X, was further investigated and was found to support the production of mature gametocytes. Infection prevalence and oocyst intensity did not differ significantly between mosquitoes given a membrane feed containing gametocytes grown in medium with 0.5% AlbuMAX + ITS-X and those grown in medium with 10% human serum. Infection prevalence and oocyst intensity was significantly higher in case of ITS–X supplementation when compared to AlbuMAX alone. Infectious gametocytes were also produced from two field clones using ITS–X supplementation. Conclusions Serum-free medium supplemented with ITS-X was able to support the growth of gametocytes of P. falciparum that were as infectious to An. gambiae as those grown in medium with 10% serum. This is the first fully serum-free culture system able to produce highly infectious gametocytes, thereby removing the requirement for access to serum for transmission assays. |
| format | Article |
| id | doaj-art-84dc876da3a8452bbd70d72cc23376f4 |
| institution | Kabale University |
| issn | 1475-2875 |
| language | English |
| publishDate | 2024-09-01 |
| publisher | BMC |
| record_format | Article |
| series | Malaria Journal |
| spelling | doaj-art-84dc876da3a8452bbd70d72cc23376f42025-08-24T11:09:13ZengBMCMalaria Journal1475-28752024-09-012311810.1186/s12936-024-05094-8Simple supplementation of serum-free medium produces gametocytes of Plasmodium falciparum that transmit to mosquitoesSabyasachi Pradhan0Prince Chigozirim Ubiaru1Lisa Ranford-Cartwright2School of Biodiversity, One Health and Veterinary Medicine, College of Medical, Veterinary and Life Sciences, University of GlasgowSchool of Biodiversity, One Health and Veterinary Medicine, College of Medical, Veterinary and Life Sciences, University of GlasgowSchool of Biodiversity, One Health and Veterinary Medicine, College of Medical, Veterinary and Life Sciences, University of GlasgowAbstract Background Human serum is a major component of Plasmodium falciparum culture medium, and can be replaced with AlbuMAX™ II, a lipid-rich bovine serum albumin, for asexual cultures. However, gametocytes produced without serum are poorly infective to mosquitoes. Serum suffers from high cost, limited availability, and variability in quality. Methods Several commercially-available media supplements were tested for their ability to support parasite growth and production of P. falciparum (3D7) gametocytes in standard RPMI1640 medium containing 0.5% AlbuMAX. The impact on asexual growth and gametocyte production with each supplement was assessed and compared to standard RPMI1640 medium containing 10% human serum, as well as to medium containing 0.5% AlbuMAX alone. The infectivity of gametocytes produced with one supplement to Anopheles gambiae sensu stricto was assessed by standard membrane feeding assay and measuring both prevalence of infection and oocyst intensity. Results Supplementation of medium containing 0.5% AlbuMAX with five supplements did not affect asexual growth of P. falciparum, and four of the five supplements supported early gametocyte production. The supplement producing the highest number of gametocytes, ITS-X, was further investigated and was found to support the production of mature gametocytes. Infection prevalence and oocyst intensity did not differ significantly between mosquitoes given a membrane feed containing gametocytes grown in medium with 0.5% AlbuMAX + ITS-X and those grown in medium with 10% human serum. Infection prevalence and oocyst intensity was significantly higher in case of ITS–X supplementation when compared to AlbuMAX alone. Infectious gametocytes were also produced from two field clones using ITS–X supplementation. Conclusions Serum-free medium supplemented with ITS-X was able to support the growth of gametocytes of P. falciparum that were as infectious to An. gambiae as those grown in medium with 10% serum. This is the first fully serum-free culture system able to produce highly infectious gametocytes, thereby removing the requirement for access to serum for transmission assays.https://doi.org/10.1186/s12936-024-05094-8Plasmodium falciparumGametocyteAlbuMAX™SerumCultureTransmission |
| spellingShingle | Sabyasachi Pradhan Prince Chigozirim Ubiaru Lisa Ranford-Cartwright Simple supplementation of serum-free medium produces gametocytes of Plasmodium falciparum that transmit to mosquitoes Malaria Journal Plasmodium falciparum Gametocyte AlbuMAX™ Serum Culture Transmission |
| title | Simple supplementation of serum-free medium produces gametocytes of Plasmodium falciparum that transmit to mosquitoes |
| title_full | Simple supplementation of serum-free medium produces gametocytes of Plasmodium falciparum that transmit to mosquitoes |
| title_fullStr | Simple supplementation of serum-free medium produces gametocytes of Plasmodium falciparum that transmit to mosquitoes |
| title_full_unstemmed | Simple supplementation of serum-free medium produces gametocytes of Plasmodium falciparum that transmit to mosquitoes |
| title_short | Simple supplementation of serum-free medium produces gametocytes of Plasmodium falciparum that transmit to mosquitoes |
| title_sort | simple supplementation of serum free medium produces gametocytes of plasmodium falciparum that transmit to mosquitoes |
| topic | Plasmodium falciparum Gametocyte AlbuMAX™ Serum Culture Transmission |
| url | https://doi.org/10.1186/s12936-024-05094-8 |
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