Assessment of Bioreactor-Derived Human Platelet Quality, Storage Profile, Safety, and Function

生物反应器衍生的人血小板质量、储存概况、安全性和功能的评估

基本信息

批准号：
9043777
负责人：
SVEN M KARLSSON
金额：
$ 72.94万
依托单位：
PLATELET BIOGENESIS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-04-01 至 2018-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9043777
关键词：
Address Adult Age Alkenes American Architecture Arterial Injury Behavior Biogenesis Biological Markers Bioreactors Birth Blood Blood Circulation Blood Platelets Blood Vessels Bone Marrow Burn injury Cell Culture Techniques Clinical Clinical Trials Clot retraction Coagulation Process Collection Computers Cytoplasmic Granules Data Devices Dimensions Drug Kinetics Embryo Emergency Situation Engineering Equipment and supply inventories Exhibits FDA approved Freezing Funding Growth HIV Half-Life Hand Hemorrhage Hemostatic Agents Hemostatic function Hour Human Human Volunteers Immune Immunocompromised Host In Vitro Infusion procedures Label Leukocytes Licensing Life Liquid substance Malignant Neoplasms Medical Megakaryocytes Metabolic Microfluidics Modeling Morphology Mus Operative Surgical Procedures Patients Perfusion Phase Physiological Pilot Projects Platelet Count measurement Platelet Transfusion Polymers Positioning Attribute Pregnancy Production Research Residual state Rest Risk Safety Sepsis Site Small Business Innovation Research Grant Source Stem cells Sterility Structure Survival Rate Temperature Teratoma Therapeutic Thrombocytopenia Thrombus Time Transfusion Transplantation Viral Virus Virus Diseases White Blood Cell Count procedure blood vessel development cancer therapy cost design humanized mouse improved in vivo induced pluripotent stem cell macrophage man meetings mortality mouse model novel parallelization polydimethylsiloxane prevent progenitor public health relevance repaired screening septic shear stress skeletal standard of care volunteer

项目摘要

DESCRIPTION (provided by applicant): Platelet BioGenesis is developing a microfluidic bioreactor to produce human platelets at clinical scale. Platelets are the 'band-aids' of the bloodstream, responsible for clot formation and blood vessel repair. Low platelet count is a significant consequence of cancer treatment, transplant, and surgery, for which platelets are a critical first-line therapy to prevent mortality due to uncontrolled bleeding. Platelet units comprising 3x1011 platelets per 200-400 mL are at present derived exclusively from human volunteer donors, and must be stored at or above 22oC to avoid irreversible temperature-related activation/aggregation. Risk of bacterial growth during room temperature storage limits shelf life to 5 days, 2 of which are consumed by bacterial screening, and 1 by transport. As a result blood centers typically do not have more than a 1.5-day platelet inventory avail- able for transfusion, which is rapidly depleted in emergencies [1, 2]. To address this major unmet need we have developed a platelet bioreactor that reproduces key features of adult bone marrow (physiological microenvironment) to trigger new platelet production at increasing scale. To date we have shown that: (1) It is feasible to generate functional megakaryocytes and platelets from human induced pluripotent stem cells (iP-SCs, a replenishable source of progenitor cells which can be stored frozen for years)[3] and (2) our bioreactor will trigger platelet formation and improve the rate and extent of platelet production from human iPSC-derived MKs above established static culture approaches[4, 5]. This Direct-to-Phase II SBIR proposal outlines 3 specific aims to determine quality, safety, and function of bioreactor-derived platelets (bdPLTs) in humanized mouse models of thrombocytopenia, and to scale platelet production to yield a human transfusion unit: Aim 1. Assess bdPLT quality, storage profile, and safety in vitro. We will evaluate platelet structure, biomarker expression, metabolic activity, and function under FDA-approved storage conditions, and determine their safety by enumerating residual leukocyte counts, and assessing viral, septic, and teratoma risk. Aim 2. Assess the hemostatic and thrombogenic potential of bdPLT in humanized mice as well as platelet clearance and circulation time. We will evaluate the in vivo behavior of bdPLTs in a novel murine model that permits human but not mouse platelet accrual at sites of arterial injury (non-GLP pilot study). Hu- man donor platelets will serve as a physiological control. Aim 3. Scale production 3,000-fold to yield a human transfusion unit (3x1011 bdPLTs per 300 mL). We will optimize our bioreactor design for continuous media perfusion and parallelization, maximize megakaryocyte zonal distribution and trapping, and equalize shear stress exposure throughout the device.

描述（由适用提供）：血小板生物发生正在开发微流体生物反应器，以临床规模产生人类血小板。血小板是血液的“创可贴”，负责凝块形成和血管修复。血小板计数低是癌症治疗，移植和手术的重要结果，该血小板是由于不受控制的出血而导致死亡率的关键一线治疗。目前，每200-400毫升符合血小板单位的3x1011血小板仅来自人类的志愿供体，必须在22oC或以上存储，以避免不可逆的温度相关激活/聚集。室温存储期间细菌生长的风险将保质期限限制为5天，其中2天被细菌筛查消耗，而通过运输1。结果，血液中心通常不超过1.5天的血小板库存可用性 - 解决了这一主要未满足的需求，我们开发了一个血小板生物反应器，该血小板生物反应器重现了成人骨髓（生理微环境）的关键特征，以触发新的血小板生产的越来越高。迄今为止，我们已经表明：（1）可行的是从人类诱导的多能干细胞中产生功能性的巨核细胞和血小板（IP-SCS，可复制的祖细胞来源，可以冷冻多年）[3]和（2）（2）（2）我们的生物表现会触发人类的范围和范围的生产范围，并触发了人类范围的范围，该培养物的范围是水平的，该培养物的范围是水平的范围。接近[4，5]。该直接到相的II SBIR提案概述了3个特定目的，目的是确定人体化小鼠血小板减少症的人体化小鼠模型中生物反应器衍生的血小板（BDPLT）的质量，安全性和功能，并缩放血小板生产以产生人类翻译单元：AIM 1。评估BDPLT质量，储存率和安全性。我们将在FDA批准的存储条件下评估血小板结构，生物标志物表达，代谢活性和功能，并通过列举残留的白细胞计数来确定其安全性，并评估病毒，化粪池和畸胎瘤风险。 AIM 2。评估人源化小鼠中BDPLT的止血和血栓形成潜力，以及血小板清除和循环时间。我们将在一种新型的鼠模型中评估BDPLT的体内行为，该模型允许人类的动脉损伤部位（非GLP初步研究）在人体的血小板精度（无小鼠血小板精度）。 Human捐赠者血小板将作为身体控制。 AIM 3。产量3,000倍以产生人类输血单元（每300毫升3x1011 bdplts）。我们将优化生物反应器设计，以进行连续的培养基灌注和副灌注，最大的巨核细胞区域分布和捕获，并在整个设备中均衡剪切应力暴露。