{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T03:24:38Z","timestamp":1792553078104,"version":"4.5.0"},"reference-count":17,"publisher":"Cambridge University Press (CUP)","issue":"2","license":[{"start":{"date-parts":[[2009,4,6]],"date-time":"2009-04-06T00:00:00Z","timestamp":1238976000000},"content-version":"unspecified","delay-in-days":6580,"URL":"https:\/\/www.cambridge.org\/core\/terms"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Parasitology"],"published-print":{"date-parts":[[1991,4]]},"abstract":"<jats:title>SUMMARY<\/jats:title><jats:p>Bloodstream forms of<jats:italic>Trypanosoma congolense<\/jats:italic>(2 clones: ILNat3\u00b71 and IL3000, and 4 stocks: IL2079, IL2466, IL3266 and CP-81) were continuously cultivated<jats:italic>in vitro<\/jats:italic>at 34\u201336 \u00b0C in the absence of feeder cell layers, using HMI-93 medium which was modified from Iscove's modified Dulbecco's MEM (Flow Laboratories, Irvine, Scotland). The modification was done by supplementing the medium with 0\u00b705 mM bathocuproine sulphonate, 1\u00b75 mM L-cysteine, 0\u00b75 mM hypoxanthine, 0\u00b712 mM 2-mercaptoethanol, 1 mM sodium pyruvate, 0\u00b716 mM thymidine, 20% (v\/v) heat-inactivated young goat serum and 5% (v\/v) Serum Plus\u2122 (Hazleton Biologics, Lenaxa, KS, USA). Trypanosomes obtained from two different sources were used to initiate primary cultures: (1) metacyclic forms which were produced<jats:italic>in vitro<\/jats:italic>at 27 \u00b0C, and (2) bloodstream forms obtained from Balb\/c mice which had been infected with the bloodstream forms transformed<jats:italic>in vitro<\/jats:italic>from the metacyclic forms. Metacyclic forms placed in 25 cm<jats:sup>2<\/jats:sup>T-type (T-25) flasks rapidly attached to the bottom of the flasks and transformed to bloodstream forms during the initial 24 h and continued to proliferate. The bloodstream forms isolated from the infected mouse blood by means of diethylaminoethyl cellulose (DE52) column chromatography also continued to proliferate in the flasks. Cultures were maintained by replacing the medium every 24 h. Every 4\u20135 days, the attached bloodstream forms were resuspended in fresh medium by gentle pipetting and then were subcultured. The method was further simplified by initiating primary cultures directly with 10 \u03bcl of the tail blood of infected mice in 24-well culture plates and then by subcultivating either in wells or in T-25 flasks. The shortest population doubling time, 9 h, was achieved by seeding subcultures with 10<jats:sup>6<\/jats:sup>bloodstream forms\/ml. The bloodstream forms propagated in this system were morphologically similar to those seen in infected mouse blood, they were covered with a surface coat as examined by electron microscopy and they were infective to mice.<\/jats:p>","DOI":"10.1017\/s0031182000062533","type":"journal-article","created":{"date-parts":[[2009,6,5]],"date-time":"2009-06-05T14:33:51Z","timestamp":1244212431000},"page":"225-236","source":"Crossref","is-referenced-by-count":106,"title":["<i>In vitro<\/i>cultivation of<i>Trypanosoma congolense<\/i>bloodstream forms in the absence of feeder cell layers"],"prefix":"10.1017","volume":"102","author":[{"given":"H.","family":"Hirumi","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"K.","family":"Hirumi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"56","published-online":{"date-parts":[[2009,4,6]]},"reference":[{"key":"S0031182000062533_ref003","doi-asserted-by":"publisher","DOI":"10.1084\/jem.162.4.1256"},{"key":"S0031182000062533_ref004","doi-asserted-by":"publisher","DOI":"10.1021\/bi00439a015"},{"key":"S0031182000062533_ref015","doi-asserted-by":"crossref","first-page":"735","DOI":"10.1128\/IAI.43.2.735-738.1984","article-title":"Resistance of cattle to tsetse-transmitted challenge with Trypanosoma brucei or Trypanosoma congolense after spontaneous recovery from syringe-passaged infections","volume":"43","author":"Nantulya","year":"1984","journal-title":"Infection and Immunity"},{"key":"S0031182000062533_ref009","doi-asserted-by":"publisher","DOI":"10.1016\/B978-0-12-427150-0.50060-7"},{"key":"S0031182000062533_ref011","doi-asserted-by":"publisher","DOI":"10.2307\/3282883"},{"key":"S0031182000062533_ref010","doi-asserted-by":"publisher","DOI":"10.1080\/00034983.1984.11811824"},{"key":"S0031182000062533_ref001","doi-asserted-by":"crossref","first-page":"1273","DOI":"10.1002\/j.1460-2075.1985.tb03772.x","article-title":"Cultivation in a semi-defined medium of animal infective forms of Trypanosoma brucei, T. equiperdum, T. evansi, T. rhodesiense and T. gambiense","volume":"4","author":"Baltz","year":"1985","journal-title":"EMBO Journal"},{"key":"S0031182000062533_ref005","first-page":"12","article-title":"Sleeping sickness survey in the Serengeti area (Tanzania) 1971. Part I. Examination of large mammals for trypanosomes","volume":"30","author":"Geigy","year":"1973","journal-title":"Acta Tropica"},{"key":"S0031182000062533_ref016","doi-asserted-by":"publisher","DOI":"10.1016\/0014-4894(74)90107-6"},{"key":"S0031182000062533_ref002","first-page":"94","volume-title":"In Vitro Methods for Parasite Cultivation","author":"Brun","year":"1987"},{"key":"S0031182000062533_ref006","doi-asserted-by":"publisher","DOI":"10.1017\/S0031182000041883"},{"key":"S0031182000062533_ref007","first-page":"118","volume-title":"In Vitro Methods for Parasite Cultivation","author":"Gray","year":"1987"},{"key":"S0031182000062533_ref008","doi-asserted-by":"publisher","DOI":"10.1016\/0166-6851(90)90075-W"},{"key":"S0031182000062533_ref012","doi-asserted-by":"publisher","DOI":"10.1126\/science.558652"},{"key":"S0031182000062533_ref013","first-page":"165","volume-title":"Tropical Diseases Research, Series 3","author":"Hirumi","year":"1980"},{"key":"S0031182000062533_ref014","doi-asserted-by":"publisher","DOI":"10.1016\/0014-4894(70)90120-7"},{"key":"S0031182000062533_ref017","doi-asserted-by":"publisher","DOI":"10.1007\/BF00930828"}],"container-title":["Parasitology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.cambridge.org\/core\/services\/aop-cambridge-core\/content\/view\/S0031182000062533","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2020,5,19]],"date-time":"2020-05-19T06:47:46Z","timestamp":1589870866000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.cambridge.org\/core\/product\/identifier\/S0031182000062533\/type\/journal_article"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1991,4]]},"references-count":17,"journal-issue":{"issue":"2","published-print":{"date-parts":[[1991,4]]}},"alternative-id":["S0031182000062533"],"URL":"https:\/\/doi.org\/10.1017\/s0031182000062533","relation":{},"ISSN":["0031-1820","1469-8161"],"issn-type":[{"value":"0031-1820","type":"print"},{"value":"1469-8161","type":"electronic"}],"subject":[],"published":{"date-parts":[[1991,4]]}}}