{"id":866,"date":"2013-09-13T14:31:29","date_gmt":"2013-09-13T13:31:29","guid":{"rendered":"http:\/\/blogs.lshtm.ac.uk\/alsfordlab\/?page_id=866"},"modified":"2014-11-26T11:30:03","modified_gmt":"2014-11-26T10:30:03","slug":"bloodstream-form-culture","status":"publish","type":"page","link":"https:\/\/blogs.lshtm.ac.uk\/alsfordlab\/protocols\/bloodstream-form-culture\/","title":{"rendered":"Bloodstream-form culture"},"content":{"rendered":"

Adapted from Hirumi & Hirumi (1994) Parasitology Today<\/em> 10<\/strong>:80<\/a><\/p>\n

HMI-9 powder is custom made by Life Technologies and is generally purchased on a consortium basis<\/p>\n

One pot is sufficient to make up 5 litres after addition of the following:<\/p>\n\n\n\n\n\n
Sodium bicarbonate<\/span><\/td>\n15 g<\/span><\/td>\n<\/tr>\n
Pen-Strep (Life Technologies<\/a>)<\/span><\/td>\n50 ml<\/span><\/td>\n<\/tr>\n
\u00df-mercaptoethanol<\/span><\/td>\n70 \u00b5l<\/span><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

Make up to 4.5 litres with distilled water, filter sterilise, store at 4\u00b0C in 450 ml volumes.<\/p>\n

Prior to use, add 50 ml fetal bovine serum (FBS; Sigma<\/a>), giving a final concentration of 10% FBS and store at 4\u00b0C for up to two months.<\/p>\n

NB: Some early publications indicate that FBS should be heat inactivated at 56\u00b0C for 30 minutes prior to use in bloodstream-form<\/em> T. brucei culture<\/em> – this is not<\/strong> necessary; in fact our unpublished data suggests that using non-inactivated FBS leads to faster, more robust cell growth.<\/em><\/p>\n

Cultures should be grown in 5% CO2<\/sub> incubator at 37\u00b0C, and should not exceed a density of 2.5×106<\/sup> cells per ml.<\/p>\n","protected":false},"excerpt":{"rendered":"

Adapted from Hirumi & Hirumi (1994) Parasitology Today 10:80 HMI-9 powder is custom made by…<\/p>\n","protected":false},"author":116,"featured_media":0,"parent":17,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"footnotes":""},"class_list":["post-866","page","type-page","status-publish","hentry","odd"],"yoast_head":"\nBloodstream-form culture - Alsford Lab<\/title>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/blogs.lshtm.ac.uk\/alsfordlab\/protocols\/bloodstream-form-culture\/\" \/>\n<meta property=\"og:locale\" content=\"en_GB\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Bloodstream-form culture - Alsford Lab\" \/>\n<meta property=\"og:description\" content=\"Adapted from Hirumi & Hirumi (1994) Parasitology Today 10:80 HMI-9 powder is custom made by...\" \/>\n<meta property=\"og:url\" content=\"https:\/\/blogs.lshtm.ac.uk\/alsfordlab\/protocols\/bloodstream-form-culture\/\" \/>\n<meta property=\"og:site_name\" content=\"Alsford Lab\" \/>\n<meta property=\"article:modified_time\" content=\"2014-11-26T10:30:03+00:00\" \/>\n<meta name=\"twitter:label1\" content=\"Estimated reading time\" \/>\n\t<meta name=\"twitter:data1\" content=\"1 minute\" \/>\n<script type=\"application\/ld+json\" class=\"yoast-schema-graph\">{\"@context\":\"https:\/\/schema.org\",\"@graph\":[{\"@type\":\"WebPage\",\"@id\":\"https:\/\/blogs.lshtm.ac.uk\/alsfordlab\/protocols\/bloodstream-form-culture\/\",\"url\":\"https:\/\/blogs.lshtm.ac.uk\/alsfordlab\/protocols\/bloodstream-form-culture\/\",\"name\":\"Bloodstream-form culture - 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