Enhanced Bioprocessing Strategies for Human Mesenchymal Stem Cells

描述(由申请人提供)： 干细胞早期临床试验的结果显然为解决各种医学问题提供了一条新的途径，这些问题包括为自体移植创造工程组织，以及修复各种器官。间充质干细胞(MSCs)具有向靶组织归巢的特性，具有分化为多种不同细胞类型的能力，并可作为旁分泌来源，从而促进邻近细胞的修复和生长，因此是干细胞治疗的候选细胞。然而，MSC治疗的一个值得注意的问题是，分离、加工和输血时会发生细胞死亡(&gt；50%)，这是有充分证据的。这一问题导致许多小组在输血前通过短暂暴露于应激状态(如低氧)来“启动”MSC。CPSI-Biotech目前正在开发一系列培养试剂CellGuard，旨在防止细胞在生物处理过程中丧失细胞活性和功能。这个第一阶段的项目旨在开发CellGuard-MSC-一种介质补充剂，旨在保护MSC在常温下的分离、处理和输血过程中，使细胞数量和分化潜力保持在更自然的状态。CellGuard是特定于细胞和过程的，基于对人体细胞在生物处理过程中激活的关键细胞应激途径的识别和调制而设计的。本提案中概述的研究将(1)确定在MSC处理过程中激活哪些应激途径，(2)确定这些途径的调节器，以及(3)测试CellGuard-MSC原型在代表生物处理和输血的条件下维持MSCs的能力。第二阶段将侧重于细胞治疗中使用的其他干细胞类型，以及CellGuard-MSC的进一步开发和特征。来源特定的MSCs，如脂肪来源的、角膜来源的和骨髓来源的MSCs将被包括在第二阶段计划中。该项目的成功完成将通过开发一种分子“盔甲”，在细胞分离和处理过程中保护骨髓间充质干细胞的完整性和分化潜力，从而加强骨髓间充质干细胞的治疗。 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类细胞。各种类型的人类细胞现在正被用于治疗癌症和糖尿病，也被用作生物医学领域中工程组织的构建块。操纵人类细胞以使其能够用于这些目的往往会损害细胞功能和生存能力。CellGuard产品组合旨在当这些细胞在临床应用的道路上受到这些恶劣条件的影响时保护它们。这一特殊项目旨在开发一套保护人类间充质干细胞的解决方案，目前正被用于从心脏细胞疗法到治疗糖尿病的各种干细胞应用。

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.