Enhanced Bioprocessing Strategies for Human Mesenchymal Stem Cells

描述（由申请人提供）： 使用干细胞的早期临床试验的结果清楚地提供了一种新的途径，以解决各种医疗问题，从创建用于自体移植的工程组织到修复各种器官。间充质干细胞（MSC）是干细胞治疗的特别有吸引力的候选者，因为它们具有归巢到靶组织的特性，分化成不同细胞类型的宿主的能力，以及作为旁分泌来源的作用，从而促进邻近细胞的修复和生长。然而，MSC疗法的一个值得注意的问题是在分离、处理和输血时发生的有据可查的细胞死亡（> 50%）。这个问题已经导致许多研究小组在输血前通过短暂暴露于应激状态如缺氧来“预处理”MSC。CPSI-Biotech目前正在开发一系列培养试剂CellGuard，旨在防止生物加工过程中人类细胞的细胞活力和功能丧失。该第1阶段项目旨在开发CellGuard-MSC -一种培养基补充剂，旨在常温下分离、处理和输血期间保护MSC，使细胞数量和分化潜力保持在更天然的状态。CellGuard是细胞和工艺特异性的，其设计基于对生物加工过程中人类细胞中激活的关键细胞应激途径的识别和调节。该提案中概述的研究将（1）确定MSC处理过程中激活的应激途径，（2）确定这些途径的调节剂，（3）测试原型CellGuard-MSC在生物处理和输血条件下维持MSC的能力。第二阶段将专注于细胞治疗中使用的其他干细胞类型，以及CellGuard-MSC的进一步开发和表征。来源特异性MSC，如脂肪来源的、角膜来源的和骨髓来源的MSC将被纳入2期计划。该项目的成功完成将通过开发一种分子“盔甲”来增强MSC细胞治疗，该分子“盔甲”将在细胞分离和处理过程中保护MSC的完整性和分化潜力。 公共卫生相关性： 该项目旨在进行研究，以开发一系列名为CellGuard的产品，该产品将在操作过程中保护人体细胞。各种人类细胞类型现在被用于治疗癌症，糖尿病，并作为生物医学领域中使用的工程组织的构建块。操纵人类细胞以使其可用于这些目的通常会损害细胞功能和活力。CellGuard产品组合旨在保护这些细胞，当它们在临床应用的道路上受到这些恶劣条件的影响时。这个特殊的项目旨在开发一套解决方案，以保护目前用于各种干细胞应用的人类间充质干细胞，从心脏细胞治疗到治疗糖尿病。

DESCRIPTION (provided by applicant): The results of early clinical trials using stem cells clearly offer the promise of a new path to addressing a variety of medical problems that range from creating engineered tissues for autologous grafts to repairing a variety of organs. Mesenchymal stem cells (MSCs) are especially attractive candidates for stem cell therapy given their documented characteristic of homing to the target tissue, ability to differentiate into a host of different cell types, and role as a paracrine source thus facilitating the repair and growth of neighboring cells. A notable problem with MSC therapy, however, is the well documented cell death (> 50%) that occurs upon isolation, processing and transfusion. This issue has led many groups to "prime" MSC prior to transfusion by transient exposure to stress regimes such as hypoxia. CPSI-Biotech is currently developing a series of culture reagents, CellGuard, that are designed to prevent the loss of cell viability and function of human cells during bioprocessing. This Phase 1 project is designed to develop CellGuard-MSC - a media supplement designed to protect MSC during isolation, processing and transfusion at normothermic temperatures so that both the cell number and differentiation potential are maintained in a more native state. CellGuard is cell and process specific and designed based on the identification and modulation of the key cell stress pathways that are activated in human cells during bioprocessing. Research outlined in this proposal will (1) determine which stress pathways are activated during MSC processing, (2) identify modulators of these pathways and (3) test a prototype CellGuard-MSC for its ability to maintain MSCs under conditions representative of bioprocessing and transfusion. Phase 2 will be focused on additional stem cell types used in cell therapy as well as further development and characterization of CellGuard-MSC. Source-specific MSCs such as adipose-derived, corneal-derived and bone marrow-derived MSCs will be included in the Phase 2 program. The successful completion of this project will result in an enhancement of MSC cell therapy through the development of a molecular "coat of armor" that will protect the integrity and differentiation potential of MSCs during cell isolation and processing. PUBLIC HEALTH RELEVANCE: This project is designed to conduct research that will lead to the development of a series of products, termed CellGuard, which will protect human cells during manipulation. A variety of human cell types are now being used to treat cancer, diabetes, and also serve as the building blocks of engineered tissues used in the biomedical field. Manipulating human cells such that they can be used for these purposes often compromises both cell function and viability. The CellGuard portfolio of products is designed to protect these cells when they are subjected to these harsh conditions on the path for clinical application. This particular project is designed to develop a set of solutions that will protect human mesenchymal stem cells that are currently being used in a variety of stem cell applications ranging from cell therapy for the heart to treating diabetes.