Continuous in vitro culture of Babesia divergens in a serum-free medium
| Autor: | Eric Precigout, Henri J. Vial, K. Moubri, André Gorenflot, J. L. Lemesre, Marie-Laure Ancelin, N. Grande, B. Carcy |
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| Rok vydání: | 1997 |
| Předmět: |
Serum albumin
Babesia Biology Culture Media Serum-Free Microbiology Andrology chemistry.chemical_compound Animals Humans Bovine serum albumin Babesia divergens HEPES Fatty Acids Albumin Serum Albumin Bovine Vitamins Glutathione biology.organism_classification Lipids In vitro Chemically defined medium Infectious Diseases chemistry biology.protein Animal Science and Zoology Parasitology |
| Zdroj: | Parasitology. 115:81-89 |
| ISSN: | 1469-8161 0031-1820 |
| DOI: | 10.1017/s0031182097008937 |
| Popis: | Babesia divergens was cultivated in RPMI 1640 (25 mM HEPES) supplemented with 10% human serum (RPMI-10% HS) with a high percentage of parasitized erythrocytes (PPE) ([ges ]40%). Standardization of in vitro tests, purification of exoantigens, biochemical studies and the safety of the culture handler motivated the development of a serum-free defined medium. Removal of serum greatly reduced the PPE but, after a period of adaptation, the culture was continuous and the parasite was able to develop a 3% routine PPE. Addition of vitamins or reduced glutathione in basal medium (RPMI) did not improve the PPE. The supplementation of basal medium with lipidic carrier (Albumax I or bovine serum albumin–Cohn's fraction V) promoted the growth of B. divergens with high PPE (>30%) close to those obtained in RPMI–10% HS. Neither protein nor lipid fractions alone were able to restore the growth of B. divergens. Nevertheless, the whole lipid fraction from serum or Albumax I added to delipidated albumin partially restored the growth (7% PPE), indicating that the presentation of specific lipids by a carrier is crucial for the parasite. All the data indicate that Albumax I can replace human serum offering the advantages of safety, standardization for chemosensitivity tests, and exoantigen purification. |
| Databáze: | OpenAIRE |
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