IN VITRO DIFFERENTIATION OF SKIN PROGENITOR CELLS: CHANGES IN GENE

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 皮肤作为细胞替代疗法的来源，提供了强大的优势，如可获得性和固有的器官再生潜力。皮肤作为体细胞干细胞的自体来源可以绕过与异种细胞替代疗法相关的典型的免疫原性排斥反应。皮肤来源的祖细胞(SKP)在体外分化为神经和神经胶质细胞。关于血统承诺的决定机制，人们知之甚少。众所周知，在体外分化时，会产生更高比例的神经胶质细胞。在小鼠和大鼠中枢神经系统(CNS)的神经前体细胞中，基因组DNA的甲基化变化控制着谱系的确定和分化的开始。这项拟议工作的工作假设是，SKP基因组DNA中诱导的低甲基化将使其基因和蛋白质表达谱向神经元谱系转移。体内和体外证据表明，DNA甲基转移酶的抑制改变了胶质-星形细胞谱系的开始和决定。使用DNA甲基转移酶抑制剂5-氮杂脱氧胞苷作为药理学工具，我们打算去甲基化褐家鼠新生儿SKP的基因组DNA。我们预计，体外去甲基化将诱导更高比例的SKP来源的细胞分化为神经元。通过确定多个神经基因表达和蛋白标记物的变化来探索神经元表型的变化。离体 SKP细胞分化的调控为进一步研究表观遗传机制如何控制细胞分化提供了一个很好的机会。了解表观遗传机制在细胞分化中的作用也有助于优化 在体外条件下，使用皮肤作为神经细胞的自体来源，用于神经疾病的细胞替代治疗。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Skin, as a source for cell replacement therapies, offers formidable advantages such as accessibility and innate organ regenerative potential. Skin as an autologous source of somatic stem cells can circumvent the quintessential immunogenic rejection associated to heterologous cell replacement therapies. In vitro differentiation of skin derived progenitor (SKP) cells produce cells of neural and glial lineages. Little is known about the mechanisms by which the decision of lineage commitment is made. It is known that upon in vitro differentiation, higher proportions of glial cells are produced. In neural progenitor cells from the central nervous system (CNS) of mouse and rats the lineage determination and onset of differentiation is controlled by methylation changes of genomic DNA. The working hypothesis of the proposed work is that an induced hypo-methylation in the genomic DNA of the SKP will shift their gene and protein expression profile towards a neuronal lineage. In vivo and in vitro evidence suggests that inhibition of DNA methyltransferases alters the onset and determination of.the glial-astrocytic lineage. With the use of a DNA methyltransferases inhibitor 5-aza-deoxycytidine as a pharmacological tool, we intend to de-methylate the genomic DNA of the SKP from Rattus norvegicus neonates. We expect that the in vitro de-methylation will induce a higher proportion of SKP derived cells to differentiate into neurons. The change in phenotype towards neurons will be explored by ascertaining the changes observed in multiple neural gene expression and protein markers. In vitro regulation of SKP cell differentiation offers a good opportunity to further study how epigenetic mechanisms control cell differentiation. Understanding the role of epigenetic mechanisms on cell differentiation also allows for optimization of in vitro conditions to use the skin as an autologous source of neurons for cell replacement therapy of neurological disorders.