ENDOTHELIAL & KERATINOCYTE STEM CELLS IN WOUND HEALING

内皮细胞

• 批准号: 6381996
• 负责人: GINA C SCHATTEMAN
• 金额: $ 25.73万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-30 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6381996
• 关键词: athymic mouse; cell population study; cell transplantation; diabetes mellitus; hematopoietic stem cells; histology; human tissue; immunocytochemistry; keratinocyte; morphometry; nonhuman therapy evaluation; pathologic process; skin; stem cells; tissue mosaicism; wound healing

In the past few years interest in stem cells as potential therapeutic agents has exploded, due in part to the recognition that stem cells are present in a myriad of tissues in the adult, and that these adult somatic stem cell populations are pluripotent. We have been studying two stem cell populations, blood-derived and keratinocyte stem cells. In Aim I we will examine the extent to which blood-derived cells are involved in wound healing by determining the contribution Of endogenous blood-derived cells to injured and uninjured skin in non-diabetic and diabetic mice. This will be accomplished by making skin wounds in hematopoietic chimeric mice (mice whose endogenous hematopoietic system has been replaced with cells that can be distinguished from those of the host). Wounds will be evaluated using histology, immunohistochemistry, and morphometry. Our hypothesis is that diabetes reduces the normal contribution of blood- derived cells to structures in the skin, especially during wound healing. If this hypothesis is correct, additional experiments in this aim will begin to delineate the mechanisms responsible for the reduction. In the second part of this proposal we will test whether blood-derived or keratinocyte stem cells can be used therapeutically to augment wound healing in a mouse model of diabetes. For this we will test whether addition of subsets of blood cells or epidermal stem cells accelerate wound healing in non-diabetic and diabetic mice. Our hypotheses are that addition of exogenous blood-derived or keratinocyte stem cells can accelerate wound healing, and that this effect will be more pronounced in diabetic than in non-diabetic mice. We also hypothesize that both blood-derived and keratinocyte stem cell populations are multipotent and will contribute to a variety of cell types in the wounds. Dil labeled leukocytes, DiO labeled skin keratinocyte stem cells, or both will be injected directly into the wound beneath the scabs of skin wounds in non-diabetic and diabetic nude mice. At various times thereafter, wounds will be harvested and examined histologically and immunohistochemically to assess the rate of wound healing. We will also investigate whether diabetes affects the incorporation of these labeled cells into different skin components.

在过去的几年中，对干细胞作为潜在治疗剂的兴趣已经激增，部分原因是认识到干细胞存在于成人的无数组织中，并且这些成人体干细胞群体是多能的。我们一直在研究两个干细胞群体，血液来源的干细胞和角质形成细胞。在目的I中，我们将通过确定内源性血液来源的细胞对非糖尿病小鼠和糖尿病小鼠中受损和未受损皮肤的贡献来检查血液来源的细胞参与伤口愈合的程度。这将通过在造血嵌合小鼠（其内源性造血系统已被可与宿主细胞区分开的细胞替代的小鼠）中制造皮肤伤口来实现。 将使用组织学、免疫组织化学和形态测定法评价伤口。我们的假设是糖尿病减少了血液来源的细胞对皮肤结构的正常贡献，特别是在伤口愈合期间。如果这一假设是正确的，在这方面的其他实验将开始描绘的机制负责减少。在本提案的第二部分，我们将测试血液来源的干细胞或角质形成细胞干细胞是否可以在糖尿病小鼠模型中用于治疗以增强伤口愈合。为此，我们将测试添加血细胞或表皮干细胞的亚群是否加速非糖尿病小鼠和糖尿病小鼠的伤口愈合。 我们的假设是，添加外源性血液来源的或角质形成细胞干细胞可以加速伤口愈合，并且这种效果在糖尿病小鼠中比在非糖尿病小鼠中更明显。我们还假设血液来源的干细胞和角质形成细胞群都是多能的，并将有助于伤口中的各种细胞类型。将Dil标记的白细胞、DiO标记的皮肤角质形成细胞干细胞或两者直接注射到非糖尿病和糖尿病裸鼠的皮肤伤口的痂下的伤口中。在此后的不同时间，将采集伤口并进行组织学和生物化学检查，以评估伤口愈合率。我们还将研究糖尿病是否会影响这些标记细胞进入不同的皮肤成分。