Analysis of the Developmental Potential of USSC in vitro and in vivo

USSCs在共培养中成功地向心肌分化，在体内，USSCs移植到梗死大鼠心脏显示出改善心功能的作用，尽管随着时间的推移移植细胞显著减少。我们现在的目标是明确USSC介导的心脏修复的细胞和分子机制(S)，并通过采取以下步骤来提高移植USSC的质量。首先，通过差异蛋白质组(DGE)和细胞因子/趋化因子(BioPlex)分析心脏间质液(在“反向心脏”模型中获得)，以确定USSC处理心脏释放的潜在生物有效旁分泌因子。其次，利用流式细胞仪和体内~(19)F-MRI定量评价炎症细胞(CD45~+、CD11b~+、CD3~+)和驻留心肌干细胞(c-kit~+、ist1~+)在心肌梗死中心区、边缘区和远区的作用。第三，SOX17、TBX5和MEF2C基因在USSCs中的基因过表达将产生心脏承诺细胞，并将通过共培养和体内实验来研究其功能相关性，以确定这是否会提高USSCs移植后的存活和整合。

Cardiomyogenic differentiation of USSCs was successfully demonstrated in co-culture, and invivo, USSCs transplanted into the infracted rat heart was shown to improve cardiac function despite significant loss of transplanted cells over time. We now aim to define the cellular and molecular mechanism(s) underlying USSC-mediated cardiac repair and to improve the quality of transplanted USSC by taking the following steps. Firstly, heart interstitial fluid (obtained in “reversed heart” model) will be subject to differential proteomic (DIGE) and cytokine/chemokine (Bioplex) analysis to define potential bio-effective paracrine factors released from USSC-treated hearts. Secondly, the role of inflammatory (CD45+, CD11b+, CD3+) and resident cardiac stem cells (c-kit+, ist1+) will be quantitatively assessed in center, border and remote zone of the infarct heart by FACS analysis as well as by using of 19F-MRI in vivo. Thirdly, cardiac committed cell will be generated by genetic overexpression of SOX17, TBX5 and MEF2c in USSCs and the functional relevance will be investigated in co-culture and in-vivo experiments to determine whether this will enhance the survival and integration of USSCs after transplantation.