DEVELOPMENT OF A NON-INVASIVE STRATEGY ENHANCING HAEMATOPOIETIC STEM CELL (HSC) POTENTIAL OF HUMAN UMBILICAL CORD CELLS

含有血液干细胞的骨髓移植称为造血干细胞（HSC）已被用于有效治疗各种血液疾病几十年。然而，由于缺乏合适的供体，这种治疗的使用受到限制。寻找高效生产HSC或富集HSC的替代来源的方法是当前血液学的关键目标之一。脐带血（UC）是HSC的一种容易获得且潜在地非常重要的来源，但其中所含的HSC的数量较低，这限制了其用于成人治疗的用途。该项目的目的是在我们实验室先前开发的基础上开发一种方法，该方法将显着增强脐带的HSC活性。HSC在胚胎发育过程中有效地产生，我们以前已经在发育中的胚胎中确定了一个强大的HSC来源，现在称为AGM区域。我们已经开发了一种新的技术，使我们能够从培养的AGM区域产生大量的HSC。在这里，我们建议使用AGM细胞增强脐带的HSC活性。我们将来自AGM区域的细胞与人脐带细胞的细胞群共培养。为了验证这种方法，并使其与临床环境兼容，我们将从人AGM区域获得永生化细胞系，并在脐带细胞上测试其诱导能力。我们将通过金标准试验评估脐带造血干细胞的产生，该试验使用移植，其中将在受体动物中评估人血细胞的长期植入。

Transplantations of bone marrow containing blood stem cells termed haematopoietic stem cells (HSC) have been used for several decades for efficient treatment of various blood diseases. However, due to shortage of appropriate donors, the use of this treatment is limited. Search for methods of efficient production of HSCs or alternative sources enriched for HSCs is one of the key goals of current haematology. Umbilical cord (UC) blood is an easily accessible and potentially very important source of HSCs but the number of HSCs contained there is low, which limits its use for treatment of adults. The aim of this project is to develop a methodology based on previous development in our laboratory which would allow significant enhancement of HSC activity of the umbilical cord. HSCs are efficiently generated during embryonic development and we have previously identified a powerful source of HSCs in the developing embryo, now termed the AGM region. We have developed a novel technique, which allowed us to generate large numbers of HSCs from the AGM region in culture. Here we propose to use the AGM cells for enhancement of the HSC activity of the umbilical cord. We will co-culture cells from the AGM region with cell populations of human umbilical cord cells. To standardise this approach and also to make it compatible with clinical settings we will derive immortalised cell lines from the human AGM region and test their inductive capacity on umbilical cord cells. We will assess the production of HSCs from the umbilical cord by the gold standard test using transplantations in which long-term engraftment of human blood cells will be assessed in recipient animals.