{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T11:24:29Z","timestamp":1789989869846,"version":"4.0.1"},"reference-count":28,"publisher":"National Academy of Sciences","issue":"13","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[1997,6,24]]},"abstract":"<jats:p>\n                    The difficulty in controlling\n                    <jats:italic>Plasmodium vivax<\/jats:italic>\n                    , the most common cause of human malaria, has been complicated by growing drug resistance. We have established a method to cycle parasite generations in continuous culture using human blood cells. Chesson strain parasites were passaged from owl monkey erythrocytes to human reticulocytes in McCoy\u2019s 5A medium modified with L-glutamine with 25 mM Hepes buffer supplemented with 20% AB\n                    <jats:sup>+<\/jats:sup>\n                    human serum. Reticulocytes were separated by differential centrifugation in homologous plasma from the peripheral blood of a hemochromatosis patient. Parasites were grown during each 48-hr cycle in a static candle jar environment until the beginning of schizogony, at about 36\u201340 hr, when reticulocytes were added and cultures transferred to a shaker for 10\u201312 hr. The addition of a concentration of 10% reticulocytes resulted in stabilizing parasite densities between 0.28 and 0.57 after cycle 3 and increasing the total number of parasites at least 2-fold with each generational cycle. Cultured parasites successfully infected an owl monkey. The morphology of cultured parasites was typical of\n                    <jats:italic>P. vivax<\/jats:italic>\n                    , with highly ameboid trophozoites evident; however, infected erythrocytes were enlarged and distorted on thin film preparations. The species identity of cultivated parasites was confirmed by analysis of the A and C 18S rRNA genes from genomic DNA and expression of only the A gene during erythrocytic asexual growth. The ability to culture\n                    <jats:italic>P. vivax<\/jats:italic>\n                    opens new opportunities to develop vaccines, test drugs, and clone parasites for genome sequencing.\n                  <\/jats:p>","DOI":"10.1073\/pnas.94.13.6786","type":"journal-article","created":{"date-parts":[[2002,7,26]],"date-time":"2002-07-26T10:31:39Z","timestamp":1027679499000},"page":"6786-6791","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":113,"title":["Continuous\n                    <i>in vitro<\/i>\n                    propagation of the malaria parasite\n                    <i>Plasmodium\u2009vivax<\/i>"],"prefix":"10.1073","volume":"94","author":[{"given":"Claudia F.","family":"Golenda","sequence":"first","affiliation":[{"name":"Department of Entomology, Walter Reed Army Institute of Research, Washington, DC 20307-5100; and Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892-0425"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jun","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Entomology, Walter Reed Army Institute of Research, Washington, DC 20307-5100; and Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892-0425"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ronald","family":"Rosenberg","sequence":"additional","affiliation":[{"name":"Department of Entomology, Walter Reed Army Institute of Research, Washington, DC 20307-5100; and Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892-0425"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[1997,6,24]]},"reference":[{"key":"e_1_3_3_1_2","first-page":"309","volume":"42","author":"Anonymous","year":"1994","unstructured":"Anonymous WHO Weekly Epidemiol Rec 42, 309\u2013314 (1994).","journal-title":"WHO Weekly Epidemiol Rec"},{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(89)91792-3"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/0035-9203(93)90294-Z"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0035-9203(96)90526-X"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.781840"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.6773146"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.7233207"},{"key":"e_1_3_3_8_2","first-page":"347","volume":"18","author":"Kitchen S F","year":"1939","unstructured":"S F Kitchen Am J Trop Med 18, 347\u2013359 (1939).","journal-title":"Am J Trop Med"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM197608052950602"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.7689250"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(92)90642-P"},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/0014-4894(88)90089-6"},{"key":"e_1_3_3_13_2","doi-asserted-by":"publisher","DOI":"10.1007\/BF00931524"},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.101.2624.377.a"},{"key":"e_1_3_3_15_2","doi-asserted-by":"publisher","DOI":"10.1038\/2241126a0"},{"key":"e_1_3_3_16_2","unstructured":"National Institutes of Health (1985) Guide for the Care and Use of Laboratory Animals NIH Publication 86\u201323 (Natl. 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