{"id":107374,"date":"2021-07-02T11:12:03","date_gmt":"2021-07-02T15:12:03","guid":{"rendered":"https:\/\/med.virginia.edu\/cell-biology\/?p=107374"},"modified":"2026-05-21T12:23:40","modified_gmt":"2026-05-21T16:23:40","slug":"directed-differentiation-of-hemogenic-endothelial-cells-from-human-pluripotent-stem-cells","status":"publish","type":"post","link":"https:\/\/med.virginia.edu\/cell-biology\/2021\/07\/02\/directed-differentiation-of-hemogenic-endothelial-cells-from-human-pluripotent-stem-cells\/","title":{"rendered":"Directed Differentiation of Hemogenic Endothelial Cells from Human Pluripotent Stem Cells"},"content":{"rendered":"<p><strong>UVA Author:<\/strong> Karen Hirschi<br \/>\n<strong>Citation:<\/strong> Nelson EA, Qiu J, Chavkin NW, Hirschi KK. Directed Differentiation of Hemogenic Endothelial Cells from Human Pluripotent Stem Cells. J Vis Exp. 2021;(169):10.3791\/62391. Published 2021 Mar 31. doi:10.3791\/62391<\/p>\n<p><strong>DOI:<\/strong><br \/>\n<strong>Pub-Med Number<\/strong>: 33871448<\/p>\n<hr \/>\n<p>Blood vessels are ubiquitously distributed within all tissues of the body and perform diverse functions. Thus, derivation of mature vascular endothelial cells, which line blood vessel lumens, from human pluripotent stem cells is crucial for a multitude of tissue engineering and regeneration applications. In vivo, primordial endothelial cells are derived from the mesodermal lineage and are specified toward specific subtypes, including arterial, venous, capillary, hemogenic, and lymphatic. Hemogenic endothelial cells are of particular interest because, during development, they give rise to hematopoietic stem and progenitor cells, which then generate all blood lineages throughout life. Thus, creating a system to generate hemogenic endothelial cells in vitro would provide an opportunity to study endothelial-to-hematopoietic transition, and may lead to ex vivo production of human blood products and reduced reliance on human donors. While several protocols exist for the derivation of progenitor and primordial endothelial cells, generation of well-characterized hemogenic endothelial cells from human stem cells has not been described. Here, a method for the derivation of hemogenic endothelial cells from human embryonic stem cells in approximately 1 week is presented: a differentiation protocol with primitive streak cells formed in response to GSK3\u03b2 inhibitor (CHIR99021), then mesoderm lineage induction mediated by bFGF, followed by primordial endothelial cell development promoted by BMP4 and VEGF-A, and finally hemogenic endothelial cell specification induced by retinoic acid. This protocol yields a well-defined population of hemogenic endothelial cells that can be used to further understand their molecular regulation and endothelial-to-hematopoietic transition, which has the potential to be applied to downstream therapeutic applications.<\/p>\n","protected":false},"excerpt":{"rendered":"<p>UVA Author: Karen Hirschi Citation: Nelson EA, Qiu J, Chavkin NW, Hirschi KK. Directed Differentiation of Hemogenic Endothelial Cells from Human Pluripotent Stem Cells. J Vis Exp. 2021;(169):10.3791\/62391. Published 2021 Mar 31. doi:10.3791\/62391 DOI: Pub-Med Number: 33871448 Blood vessels are ubiquitously distributed within all tissues of the body and perform diverse functions. Thus, derivation of [&hellip;]<\/p>\n","protected":false},"author":1740,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"inline_featured_image":false,"footnotes":"","_links_to":"","_links_to_target":""},"categories":[13],"tags":[21],"class_list":["post-107374","post","type-post","status-publish","format-standard","hentry","category-featured-publications","tag-delete"],"acf":false,"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v27.4 - https:\/\/yoast.com\/product\/yoast-seo-wordpress\/ -->\n<title>Directed Differentiation of Hemogenic Endothelial Cells from Human Pluripotent Stem Cells - Department of Cell Biology<\/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:\/\/med.virginia.edu\/cell-biology\/2021\/07\/02\/directed-differentiation-of-hemogenic-endothelial-cells-from-human-pluripotent-stem-cells\/\" \/>\n<meta property=\"og:locale\" content=\"en_US\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Directed Differentiation of Hemogenic Endothelial Cells from Human Pluripotent Stem Cells - Department of Cell Biology\" \/>\n<meta property=\"og:description\" content=\"UVA Author: Karen Hirschi Citation: Nelson EA, Qiu J, Chavkin NW, Hirschi KK. Directed Differentiation of Hemogenic Endothelial Cells from Human Pluripotent Stem Cells. J Vis Exp. 2021;(169):10.3791\/62391. Published 2021 Mar 31. doi:10.3791\/62391 DOI: Pub-Med Number: 33871448 Blood vessels are ubiquitously distributed within all tissues of the body and perform diverse functions. Thus, derivation of [&hellip;]\" \/>\n<meta property=\"og:url\" content=\"https:\/\/med.virginia.edu\/cell-biology\/2021\/07\/02\/directed-differentiation-of-hemogenic-endothelial-cells-from-human-pluripotent-stem-cells\/\" \/>\n<meta property=\"og:site_name\" content=\"Department of Cell Biology\" \/>\n<meta property=\"article:published_time\" content=\"2021-07-02T15:12:03+00:00\" \/>\n<meta property=\"article:modified_time\" content=\"2026-05-21T16:23:40+00:00\" \/>\n<meta name=\"author\" content=\"zkr7e@virginia.edu\" \/>\n<meta name=\"twitter:card\" content=\"summary_large_image\" \/>\n<meta name=\"twitter:label1\" content=\"Written by\" \/>\n\t<meta name=\"twitter:data1\" content=\"zkr7e@virginia.edu\" \/>\n\t<meta name=\"twitter:label2\" content=\"Est. reading time\" \/>\n\t<meta name=\"twitter:data2\" content=\"2 minutes\" \/>\n<script type=\"application\/ld+json\" class=\"yoast-schema-graph\">{\"@context\":\"https:\\\/\\\/schema.org\",\"@graph\":[{\"@type\":\"Article\",\"@id\":\"https:\\\/\\\/med.virginia.edu\\\/cell-biology\\\/2021\\\/07\\\/02\\\/directed-differentiation-of-hemogenic-endothelial-cells-from-human-pluripotent-stem-cells\\\/#article\",\"isPartOf\":{\"@id\":\"https:\\\/\\\/med.virginia.edu\\\/cell-biology\\\/2021\\\/07\\\/02\\\/directed-differentiation-of-hemogenic-endothelial-cells-from-human-pluripotent-stem-cells\\\/\"},\"author\":{\"name\":\"zkr7e@virginia.edu\",\"@id\":\"https:\\\/\\\/med.virginia.edu\\\/cell-biology\\\/#\\\/schema\\\/person\\\/b4929a1d4683383a392718e12166cd7f\"},\"headline\":\"Directed Differentiation of Hemogenic Endothelial Cells from Human Pluripotent Stem Cells\",\"datePublished\":\"2021-07-02T15:12:03+00:00\",\"dateModified\":\"2026-05-21T16:23:40+00:00\",\"mainEntityOfPage\":{\"@id\":\"https:\\\/\\\/med.virginia.edu\\\/cell-biology\\\/2021\\\/07\\\/02\\\/directed-differentiation-of-hemogenic-endothelial-cells-from-human-pluripotent-stem-cells\\\/\"},\"wordCount\":297,\"keywords\":[\"Delete\"],\"articleSection\":[\"Featured Publications\"],\"inLanguage\":\"en-US\"},{\"@type\":\"WebPage\",\"@id\":\"https:\\\/\\\/med.virginia.edu\\\/cell-biology\\\/2021\\\/07\\\/02\\\/directed-differentiation-of-hemogenic-endothelial-cells-from-human-pluripotent-stem-cells\\\/\",\"url\":\"https:\\\/\\\/med.virginia.edu\\\/cell-biology\\\/2021\\\/07\\\/02\\\/directed-differentiation-of-hemogenic-endothelial-cells-from-human-pluripotent-stem-cells\\\/\",\"name\":\"Directed Differentiation of Hemogenic Endothelial Cells from Human Pluripotent Stem Cells - 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