Towards Large-Scale Platelet Production from Adult Hematopoietic Stem and Progenitor Cells

该奖项是根据2009年《美国复苏和再投资法案》(公法111-5)资助的。虽然几个研究小组已经证明了在培养中从巨核细胞(MKs)中生产少量功能性血小板的能力，但使用目前的技术生产一次输血剂量所需的3-6x10-11血小板将需要2.5亿CD34+造血(血液)干细胞和祖细胞。由于采集的脐带血通常含有不到500万个CD34+细胞，而从经过G-CSF(MPB)等细胞因子治疗(或动员)的捐献者采集的外周血液中含有多达2亿个CD34+细胞，因此必须将血小板产量提高几个数量级才能实际应用，该项目的目标是能够从100万个MPB CD34+细胞生产一次血小板输注剂量-或每个输入CD34+细胞生产500,000个血小板。Broader Impact血小板输注通常用于广泛的血栓形成缺陷，在美国和欧洲每年都有数百万个单位的输注。然而，为一次输血收集足够的血小板，要么需要昂贵的分离设备，要么需要汇集4到8个不同捐赠者的血小板。还有对细菌污染、血液传播病原体和受者同种异体免疫的担忧。使用良好的制造规范从培养中的巨噬细胞中生产自体或完全匹配的血小板有可能缓解所有这些问题，但仍有许多挑战需要解决。这项拟议研究的成功完成将使培养衍生血小板的大规模生产更接近实现。该项目还将有助于培训高素质的个人。从事这个项目的研究生和本科生将在跨学科的环境中接受培训，其中包括让生命科学教师参加研究生论文委员会，以及与医学院的教师和研究生合作。研究生将与其他工程和生命科学系的学生一起积极参与，并在西北大学生物技术博士前培训项目的研讨会和期刊俱乐部展示他们的工作，该项目还提供与来自行业的科学家和工程师的互动。每名研究生每年至少参加一次科学会议，并在工程学、血液学和跨学科会议上展示他们的成果并与其他与会者互动。PIS将共同努力，为化学和生物工程系和这一项目招收代表不足的少数族裔和女性研究生，并监督了很大一部分女性和少数族裔学生。皮？S以前是博士生，现在受雇于不同的制药、生物技术和工程公司。

This award is funded under the American Recovery and Reinvestment Act of 2009 (Public Law 111-5).0853603W. MillerIntellectual merit Although several research groups have demonstrated the capability to produce small quantities of functional platelets from megakaryocytic cells (Mks) in culture, producing the 3-6 x 10-11 platelets required for a single transfusion dose using current technology would require 250 million CD34+ hematopoietic (blood) stem and progenitor cells. Since an umbilical cord blood harvest typically contains less than 5 million CD34+ cells and peripheral blood harvested from donors treated (or mobilized) with cytokines such as G-CSF (mPB) contains up to 200 million CD34+ cells, the platelet yield must be increased by several orders of magnitude to be practical, The objective of this project is to enable the production of one platelet transfusion dose from one million mPB CD34+ cells - or 500,000 platelets per input CD34+ cell.Broader impacts Platelet transfusions are routinely used for a wide range of thrombotic deficiencies, and several million units are transfused each year in the USA and Europe. However, collecting enough platelets for a single transfusion requires either expensive apheresis equipment or the pooling of platelets from 4 to 8 different donors. There are also concerns with bacterial contamination, blood-borne pathogens, and alloimmunization of recipients. Production of autologous or fully matched platelets from Mks in culture using Good Manufacturing Practices has the potential to alleviate all of these problems, but many challenges remain to be addressed. Successful completion of the proposed research would bring the large-scale production of culture-derived platelets closer to fruition. This project will also contribute to the training of highly qualified individuals. Graduate students and undergraduates working on this project will be trained in an interdisciplinary environment that includes having life science faculty members on graduate student thesis committees and collaboration with faculty and graduate students in the medical school. Graduate students will actively participate, together with students from other engineering and life science departments, and present their work in the seminars and journal club of Northwestern University's predoctoral Biotechnology Training Program, which also provides interactions with scientists and engineers from industry. Each graduate student will attend at least one scientific meeting per year and will present their results and interact with other attendees at engineering, hematology, and interdisciplinary meetings. The PIs will make a concerted effort to recruit underrepresented minority and women graduate students for the Department of Chemical and Biological Engineering and this project, and has supervised a large fraction of female and minority students. The PI?s former doctoral students are employed by diverse pharmaceutical, biotechnology, and engineering companies.