Cardiac mesenchymal progenitors from postmortem cardiac tissue retained cellular characterization. Mus musculus

Background and Objective: Currently, the cells for transplantation were derived from either autologous or allogeneic tissue. The former has a drawback that the quality of donor cells could depend on the patient’s condition, and the quantity could also be limited. To solve these problems, we investigated the potential of allogeneic cardiac mesenchymal progenitors (CMPs) derived from postmortem heart, which might be an immunological privileged like bone marrow-derived mesenchymal progenitors. Materials and Methods: We examined whether viable CMPs could be isolated from murine postmortem cardiac tissue that was harvested 24 hours postmortem. After two to three weeks propagation with high dose of basic fibroblast growth factor, we performed the cellular characteristics analyses, which included proliferation and differentiation property flow cytometric analyses, and microarray analyses. Results: Postmortem CMPs had longer lag phase after seeding than CMPs from living tissues, but they demonstrated the similar characteristics in all above examinations. In addition, global gene expression analysis by microarray indicated the similar characteristics between the cell derived from postmortem and living tissue. Conclusion: These results indicate allogeneic postmortem CMPs could have promising potential for cell transplantation as clinical applications, because of circumventing the issue of brain death. Overall design: The samples were collected fom living or postmortem cardiac tissue (24 hr at 4C). We generated cardiac mesenchymal progenitors (CMPs) from these cardiac tissue, and compared global gene expression by AgilentMouse GE 8x60k Microarray. Adult, or fetal mouse heart RNAs were used as positive control. Adult mouse total heart RNAs were purchased from Clontech. Fetal mouse heart was extracted from fetus which is embryonic day 16.5 C57BL/6 strain. Tg means Transgenic mouse (C57BL/6-Tg(Myh6-EGFP)MG2).

背景与研究目的：目前临床用于移植的细胞均源自自体或异体组织。其中自体移植的缺陷在于，供体细胞的质量依赖患者自身身体状况，且细胞数量往往受限。为解决上述问题，本研究探讨了死后心脏来源的异体心脏间充质祖细胞（cardiac mesenchymal progenitors, CMPs）的应用潜力，这类细胞或许与骨髓来源的间充质祖细胞一样具有免疫豁免特性。
材料与方法：本研究检测了能否从死后24小时采集的小鼠心脏组织中分离获得存活的CMPs。经高剂量碱性成纤维细胞生长因子（basic fibroblast growth factor）传代培养2至3周后，我们开展了细胞特性分析，包括增殖与分化特性检测、流式细胞术分析，以及基因芯片分析。
研究结果：死后来源的CMPs在接种后的滞缓期较活体组织来源的CMPs更长，但在上述所有检测中均展现出相似的细胞特性。此外，基因芯片全基因表达分析结果显示，死后来源与活体来源的CMPs具有高度相似的基因表达特征。
结论：本研究结果表明，异体死后来源的CMPs有望应用于临床细胞移植治疗，因其可规避脑死亡供体相关的限制问题。
实验整体设计：实验样本采集自活体小鼠心脏组织，或4℃保存24小时的死后小鼠心脏组织。我们从上述两类心脏组织中分离培养得到心脏间充质祖细胞（CMPs），并通过安捷伦小鼠全基因组表达8x60k基因芯片（AgilentMouse GE 8x60k Microarray）比较二者的全基因表达谱。本研究以成年或胚胎期小鼠心脏RNA作为阳性对照：成年小鼠心脏总RNA购自Clontech公司；胚胎期小鼠心脏RNA提取自胚胎发育第16.5天的C57BL/6品系胎鼠；Tg指转基因小鼠（C57BL/6-Tg(Myh6-EGFP)MG2）。