A NOVEL APPROACH TO GMP PRODUCTION, CRYOPRESERVATION AND DISTRIBUTION OF TOOTH DE

一种 GMP 生产、冷冻保存和分配牙齿的新方法

基本信息

批准号：
7935375
负责人：
ERIK J. WOODS
金额：
$ 47.18万
依托单位：
GENERAL BIOTECHNOLOGY, LLC
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-19 至 2011-07-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): The overall goal of this proposal is to build on our understanding of the collection, processing and cryobiological characteristics of stem/progenitor cells recovered from extracted teeth with the ultimate culmination of these data to develop a cryobank of dental pulp derived stem cells (DPSCs) for research and/or potential transplant or other clinical utilization as a human cell product processed under current good tissue practice (cGTP) as described under 21 CFR 1271. Recent studies investigating this exciting source of post-natal stem cells have identified a population of clonogenic and highly proliferative cells derived from enzymatically digested dental pulp tissue (Gronthos et al., 2000; Perry et al., 2008). Results from these studies have indicated that these cells may have the potential to be utilized in stem cell mediated therapies as well as tissue engineering applications (Seo et al., 2005). To allow further study and potential therapeutic use of these cells, our group has begun to evaluate the ability of these cells and their respective tissues of origin to survive cryopreservation processing. Ideally, the optimum cryopreservation process will be straight forward and effective when applied to the tissue as a whole, with the idea that stem cells could be extracted post-thaw. The rationale for this would be to preserve clinical samples for subsequent stem cell recovery as it is reasonable to speculate that cryopreservation of tissues in the clinic will be more practical than direct primary isolation of stem cells, which would require additional equipment and personnel (Seo et al., 2005; Perry et al., 2008). A cryobank of these cells would add great flexibility to their use by allowing, among other things, shipment of cells to investigators and/or clinicians and adequate time for donor characterization and/or potential testing. To achieve this overall goal, we propose the following Specific Aims: (I) Final development of optimized, cGTP protocols for development of a tooth stem cell bank for distribution and use; (II) Final development cGTP protocols for cryobanking of tooth tissue for later stem/progenitor cell isolation and expansion; and (III) Development and validation of a closed container for packaging and distribution of cells for clinical use. The ultimate goal of the proposal is a cryobank of DPSCs or tissue with e90% consistent functional recovery of cells based on the most sensitive assays available when compared to their unfrozen counterparts. For Specific Aim III the goal will be an FDA approved packaging system useful for clinical utilization of stem/progenitor cells derived from teeth. This system will also be useful for other clinically relevant stem cell sources where smaller volumes are required. PUBLIC HEALTH RELEVANCE: Stem cells can develop into different tissues types in the body. "Post-natal" or "adult" stem cells offer an alternative to obtaining stem cells from embryos. Such stem cells exist in human teeth, but more research is required before these cells can have direct clinical use. A bank of frozen stem cells from teeth would be ideal to facilitate this research. This proposed project would optimize methods for processing and freezing these cells for ease of distribution for research, testing, and eventually clinical use, as well as present the ability for one to store tissue from their own teeth for potential use of their own cells if/when needed.

描述（由申请人提供）：该提案的总体目标是建立在我们对从拔牙中回收的干细胞/祖细胞的收集、处理和冷冻生物学特征的理解的基础上，并最终根据这些数据开发牙髓冷冻库衍生干细胞 (DPSC) 用于研究和/或潜在移植或其他临床用途，作为根据 21 CFR 1271 中所述的当前良好组织实践 (cGTP) 加工的人类细胞产品。最近的研究调查了这一令人兴奋的来源出生后干细胞已鉴定出源自酶消化牙髓组织的克隆形成和高度增殖细胞群（Gronthos 等，2000；Perry 等，2008）。这些研究的结果表明，这些细胞可能具有用于干细胞介导的疗法以及组织工程应用的潜力（Seo et al., 2005）。为了进一步研究这些细胞和潜在的治疗用途，我们的小组已开始评估这些细胞及其各自的起源组织在冷冻保存过程中存活的能力。理想情况下，当应用于整个组织时，最佳的冷冻保存过程将是简单有效的，并且可以在解冻后提取干细胞。这样做的理由是保存临床样本以供随后的干细胞回收，因为可以合理地推测，在诊所中冷冻保存组织将比直接初级分离干细胞更实用，后者需要额外的设备和人员（Seo et al.等人，2005；佩里等人，2008）。这些细胞的冷冻库将为其使用增加极大的灵活性，除其他外，允许将细胞运送给研究人员和/或临床医生，并有足够的时间进行供体表征和/或潜在测试。为了实现这一总体目标，我们提出以下具体目标： (I) 最终开发优化的 cGTP 方案，以开发用于分发和使用的牙齿干细胞库； (II) 最终开发的 cGTP 方案，用于牙齿组织冷冻库，以便以后干细胞/祖细胞分离和扩增； (III) 开发和验证用于临床使用细胞的包装和分配的密闭容器。该提案的最终目标是建立一个 DPSC 或组织冷冻库，与未冷冻的对应物相比，基于可用的最灵敏的检测，其细胞功能恢复达到 90% 一致。具体目标 III 的目标是建立 FDA 批准的包装系统，可用于临床利用源自牙齿的干细胞/祖细胞。该系统也可用于需要较小体积的其他临床相关干细胞来源。公共健康相关性：干细胞可以发育成体内不同的组织类型。 “产后”或“成年”干细胞提供了从胚胎获取干细胞的另一种选择。这种干细胞存在于人类牙齿中，但在这些细胞可以直接应用于临床之前还需要更多的研究。来自牙齿的冷冻干细胞库将是促进这项研究的理想选择。该拟议项目将优化处理和冷冻这些细胞的方法，以便于研究、测试和最终临床使用的分配，并提供人们储存自己牙齿组织的能力，以便在以下情况下使用自己的细胞：当需要时。