Skin Stem Cells: Purification and Characterization

皮肤干细胞：纯化和表征

• 批准号: 6705196
• 负责人: ELAINE FUCHS
• 金额: $ 35.56万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-02-26 至 2008-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6705196
• 关键词: biomarker; cell differentiation; cell population study; cell sorting; clinical research; congenital skin disorder; fluorescent dye /probe; functional /structural genomics; genetically modified animals; human subject; human tissue; keratinocyte; laboratory mouse; patient oriented research; skin; stem cells; tissue /cell culture

DESCRIPTION (provided by applicant): Stem cells are natural units of tissue repair and homeostasis. Recent studies suggest that stem cells may be more versatile than previously appreciated. This property holds promise for tissue regeneration, particularly for less controversial adult stem cells. Our global objective is to develop strategies for the isolation, purification and characterization of multipotent adult skin stem cells and their transitamplifying progeny. While skin keratinocytes are one of the few adult stem cells that can be maintained and propagated in vitro, their identification and isolation have been hampered by the lack of reliable markers that distinguish stem cells from their progeny. Without this information, the stress of removing stem cells from their niche make it impossible to assess at what point these cells might cease to function as multipotent stem cells. Here, we propose a novel strategy for purification of multipotent adult murine skin stem cells from their natural niche, by taking advantage of the slow-cycling properties and the few known markers preferentially expressed in these cells. We also devise strategies for purifying three different transit-amplifying progeny of these cells. We will determine and compare the global expression patterns of these cell populations and employ these and known markers to examine how skin stem cells change programs of gene expression during both embryonic development, and postnatally, in natural homeostasis, with the hair cycle, in response to injury or tumorigenesis, and when placed in culture. Finally, we will use this information to explore the functional consequences of altering the expression programs of key genes, which distinguish skin stem cells from their non-stem lineages. Taken together, these studies are expected to provide new and important insights into understanding how skin stem cells possess and maintain their unique self-renewing properties, and how they adopt different fates, including epidermal and hair follicle differentiation. This research is a fundamental prerequisite to ascertaining the potential of skin stem cells for therapeutic purposes that go beyond their present use in providing long-term engraftment on burn patients.

描述（由申请人提供）：干细胞是组织修复和稳态的天然单位。最近的研究表明，干细胞可能比以前所欣赏的更通用。该特性具有组织再生的希望，特别是对于较少有争议的成年干细胞。我们的全球目标是制定策略来隔离，纯化和表征多能成人皮肤干细胞及其过度放大后代的策略。虽然皮肤角质形成细胞是可以在体外维持和传播的少数成年干细胞之一，但由于缺乏将干细胞与后代区分开的可靠标记，它们的鉴定和分离受到了阻碍。如果没有这些信息，从其利基市场中去除干细胞的压力使得无法评估这些细胞在什么时候可能不再充当多能干细胞。在这里，我们提出了一种新型的策略，以利用慢循环特性和在这些细胞中优先表达的少数已知标记来纯化其自然细胞的多能鼠皮肤干细胞。我们还制定了净化这些细胞的三个不同的扩增后代的策略。我们将确定和比较这些细胞群体的全球表达模式，并采用这些和已知的标记来检查皮肤干细胞在胚胎发育过程中如何改变基因表达程序，以及在自然稳态中，在自然稳态中与毛发周期，响应损伤或肿瘤的响应，以及放置在培养中时。最后，我们将使用这些信息来探索改变关键基因表达程序的功能后果，这些基因将皮肤干细胞与其非茎谱系区分开。综上所述，这些研究有望提供新的重要见解，以了解皮肤干细胞如何拥有并保持其独特的自我更新特性，以及它们如何采用不同的命运，包括表皮和毛囊分化。这项研究是确定皮肤干细胞的潜在治疗目的潜力的基本先决条件，这些目前超越了目前的用途，可以为烧伤患者提供长期植入。