Development of selection techniques for purifying stem cells from dental tissues

开发从牙组织中纯化干细胞的选择技术

• 批准号: 7588220
• 负责人: Shaomian Yao
• 金额: $ 7.4万
• 依托单位: LOUISIANA STATE UNIV A&M COL BATON ROUGE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-15 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7588220
• 关键词: ABCB1 gene; ABCG2 gene; ATP-Binding Cassette Transporters; Acetaldehyde; Adult; Aldehydes; Binding; Body Temperature; Bypass; Cell Culture Techniques; Cell Survival; Cells; Cessation of life; Characteristics; Chemicals; Clinical; Computer software; Data; Defect; Dental; Dental Pulp; Dental Sac; Development; Disease; Doxorubicin; Foundations; Gene Expression; Goals; Healthcare; Heat-Shock Response; Heating; Human; Hypoxia; Immune; Mammals; Medical; Membrane; Methods; Microarray Analysis; Mitoxantrone; Molecular; Natural regeneration; Organ; Oxygen; Patients; Pharmaceutical Preparations; Population; Population Heterogeneity; Proliferating; Proliferation Marker; Property; Proteins; Protocols documentation; Pump; Regenerative Medicine; Research; Signal Transduction; Stem cells; Stress; Survival Rate; System; Techniques; Testing; Tissue Engineering; Tissues; Tooth Loss; Tooth structure; United States; Wolves; adult stem cell; aldehyde dehydrogenases; base; cost effective; design; dosage; drug mechanism; human disease; killings; lipid biosynthesis; member; planetary Atmosphere; repaired; research study; response; stem cell population; stem cell technology; theories

DESCRIPTION (provided by applicant): Use of stem cells to regenerate or repair damaged/defective tissues is a promising strategy to cure many human diseases and disorders. Because the current protocols for purification of stem cells from dental tissues are labor intensive, somewhat vague and often expensive, the general objective of this proposal is to develop safe, simple and efficient techniques to purify stem cells from dental tissues. Adult stem cells have been identified in many tissues including dental pulps and dental follicles. One of the important purposes of stem cell's presence in adult tissues is to serve as the repair/regeneration system to replace damaged or defective tissues/organs. In order to fulfill the repair task, stem cells must first survive the conditions causing the damage. Thus, they are presumed to possess specific or extra defense or protection mechanisms that allow them to survive various stresses. Support for this theory includes evidence that certain stem cells are equipped with a high level of efflux proteins, such as ATP-Binding Cassette (ABC) transporters on their membrane to pump out harmful chemicals (Goodell et al., 1996; Wolf et al., 1993; Zhou et al., 2002), and that some stem cells produce high levels of cytosolic aldehyde dehydrogenases (ALDHs) to detoxify cellular aldehyde (Cai et al., 2004; Storms et al., 1999; Kastan et al., 1990). When subjected to hypoxia, stem cells grow more rapidly and form more colonies in hypoxic conditions than they do in normal oxygen atmosphere (Lennon et al., 2001) suggesting that stem cells can survive better in low oxygen stress. Based on the above, the overall hypothesis is that stem cells from dental tissues possess unique selfprotection/ defense abilities (i.e., anti-stress/drug mechanisms), and that this unique feature can be used to develop conditions that selectively kill/eliminate non-stem cells such that one could purify stem cells from heterogeneous cell populations. To test this hypothesis, heterogeneous cell populations from dental pulps and dental follicles will be subjected to medium containing selected substrates of BCRP or ALDHs. In addition, the heterogeneous populations will also be cultured under thermal and hypoxic conditions. The resulting subpopulation cells will be evaluated for stem cell properties. The optimal putative stem cell populations will be further studied for their molecular characteristics using microarray. Relevance: Dental damage (defect and loss of teeth) is a major health care issue in the United States. Stem cell technology provides a very promising strategy to repair or regenerate damaged/defective teeth. Development of safe, simple and efficient techniques to purify stem cells from dental tissues is the foundation for successfully use the strategy both in research and clinical aspects.

描述（由申请人提供）：使用干细胞再生或修复受损/缺陷组织是治疗许多人类疾病和病症的一种有希望的策略。由于目前从牙齿组织中纯化干细胞的方案是劳动密集型的，有些模糊，往往是昂贵的，这个建议的总体目标是开发安全，简单和有效的技术，从牙齿组织中纯化干细胞。成体干细胞已在包括牙髓和牙囊在内的许多组织中被鉴定。干细胞存在于成体组织中的重要目的之一是作为修复/再生系统来替换受损或有缺陷的组织/器官。为了完成修复任务，干细胞必须首先在造成损伤的条件下存活。因此，他们被认为拥有特定或额外的防御或保护机制，使他们能够在各种压力下生存。对这一理论的支持包括某些干细胞配备有高水平的外排蛋白的证据，例如在其膜上的ATP结合盒（ABC）转运蛋白，以泵出有害化学物质（Goodell等人，1996; Wolf等人，1993; Zhou等人，2002），并且一些干细胞产生高水平的细胞溶质醛脱氢酶（ALDH）以使细胞醛解毒（Cai等人，2004; Storms等人，1999; Kastan等人，1990年）。当经受缺氧时，干细胞在缺氧条件下比在正常氧气气氛中生长得更快并形成更多的集落（Lennon等人，2001年），这表明干细胞可以在低氧胁迫下更好地存活。基于上述，总体假设是来自牙组织的干细胞具有独特的自我保护/防御能力（即，抗应激/药物机制），并且这种独特的特征可用于开发选择性杀死/消除非干细胞的条件，使得可以从异质细胞群中纯化干细胞。为了检验这一假设，将来自牙髓和牙囊的异质细胞群置于含有选定BCRP或ALDH底物的培养基中。此外，异质群体也将在热和缺氧条件下培养。将评价所得亚群细胞的干细胞特性。最佳假定的干细胞群体将进一步研究其分子特征，使用微阵列。相关性：牙齿损伤（牙齿缺陷和缺失）是美国的一个主要医疗保健问题。干细胞技术为修复或再生受损/有缺陷的牙齿提供了一个非常有前途的策略。安全、简便、高效的牙源性干细胞纯化技术的开发是该策略成功应用于研究和临床的基础。