Propagation of babesia parasites
AssigneeUniversity of Illinois Foundation
InventorsMiodrag Ristic, Michael G. Levy
Life of this patent
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Abstract
A method for quantitative propagation of hemotropic parasites, especially Babesia parasites, in vitro, Babesia-infected erythrocyte cultures are incubated in an environment of enhanced carbon dioxide tension wherein the culture medium overlay provides a barrier to oxygen transmission such that cellular hemoglobin is essentially maintained in its deoxy state. By varying environmental carbon dioxide tension, merozoite stages of the parasite are selectively reversibly induced to migration between residence in host cells or culture medium, rendering possible the isolation of infective antigenic merozoites in large quantities. Specific Babesia antigen is harvested in large quantity from the in vitro culture medium.
Claims16
In the method for in vitro propagation of Babesia parasites by incubation in a culture medium containing erythrocyte host cells, the improvement comprising:
- (1) providing as a component in said culture medium from about 25 to about 55 percent defibrinated serum;
- and
- (2) carrying out said incubation step,
- (a) in the presence of a controlled atmosphere containing 3 to 7 percent carbon dioxide, and
- (b) under conditions of reduced oxygen tension upon the infected erythrocytes resulting in maintenance of erythrocytic hemoglobin in its deoxy state.
In a method for in vitro propagation of Babesia bovis parasite by incubation in a culture medium containing bovine erythrocyte host cells, the improvement comprising:
- (1) providing as a component in said culture medium from about 25 to about 55 percent defibrinated bovine serum;
- and
- (2) carrying out said incubation step,
- (a) in the presence of a controlled atmosphere containing 3 to 7 percent carbon dioxide, and
- (b) under conditions of reduced oxygen tension upon infected erythrocytes resulting in maintenance of erythrocytic hemoglobin in its deoxy state.
In the method for in vitro propagation of hemotropic parasites by incubation in a culture medium containing erythrocyte host cells, the improvement comprising:
- (1) providing as a component in said culture medium from about 25 to about 55 percent defibrinated serum;
- and
- (2) carrying out said incubation step,
- (a) statically,
- (b) in the presence of a controlled atmosphere containing 3 to 6 percent carbon dioxide, and
- (c) under conditions of reduced oxygen tension upon the infected erythrocytes resulting in maintenance of erythrocytic hemoglobin in its deoxy state.
Dependencies are inferred from the claim text.