Department of Biomedical Sciences, Research Center, Montreal Heart Institute, 5000 Belanger Street, Montreal, (Quebec) H1T 1C8, Canada.
Biomaterials. 2011 Aug;32(22):5046-55. doi: 10.1016/j.biomaterials.2011.04.002. Epub 2011 May 5.
The aim of the present study was to develop a new biopolymer to increase endothelial progenitor cells (EPC) survival and amplification. As a cell culture platform, bone marrow-derived cells (BMDC) were used to investigate the biocompatibility of chitosan-phosphorylcholine (CH-PC). On CH-PC, BMDC were found in colonies with a mortality rate similar to that of fibronectin (FN), the control matrix. Adhesion/proliferation assays demonstrated a greater number of BMDC on CH-PC after 7 days with an amplification phase occurring during the second week. Difference in adhesion mechanisms between (CH-PC) and the control FN matrix suggest distinctive cell retention ability. Confocal microscopy analyses confirmed that (CH-PC) supported the survival/differentiation of endothelial cells. Moreover, flow cytometry analyses demonstrated that, (CH-PC) increased the percentage of progenitor cells (CD117(+)CD34(+)) (7.1 ± 0.8%, FN: 4.1 ± 0.8%) and EPC (CD117(+)CD34(+)VEGFR-2(+)CD31(+)) (2.33 ± 0.6%, FN: 0.25 ± 0.1%), while the mesenchymal stem cell fraction (CD44(+)CD106(+)CD90(+)) was decreased (0.07 ± 0.01%, FN: 0.55 ± 0.22%). Polymeric substrate CH-PC might provide a suitable surface to promote the amplification of EPC for future vascular therapeutic applications.
本研究旨在开发一种新的生物聚合物,以提高内皮祖细胞 (EPC) 的存活率和扩增率。作为细胞培养平台,骨髓来源的细胞 (BMDC) 用于研究壳聚糖-磷酸胆碱 (CH-PC) 的生物相容性。在 CH-PC 上,BMDC 以类似于纤维连接蛋白 (FN) 的死亡率形成集落,FN 是对照基质。粘附/增殖测定表明,在第 7 天,CH-PC 上的 BMDC 数量更多,并且在第二周发生扩增阶段。(CH-PC)和对照 FN 基质之间的粘附机制差异表明具有不同的细胞保留能力。共聚焦显微镜分析证实 CH-PC 支持内皮细胞的存活/分化。此外,流式细胞术分析表明,CH-PC 增加了祖细胞(CD117(+)CD34(+))(7.1 ± 0.8%,FN:4.1 ± 0.8%)和 EPC(CD117(+)CD34(+)VEGFR-2(+)CD31(+))(2.33 ± 0.6%,FN:0.25 ± 0.1%)的百分比,而间充质干细胞部分(CD44(+)CD106(+)CD90(+))减少(0.07 ± 0.01%,FN:0.55 ± 0.22%)。多聚物基质 CH-PC 可能为未来血管治疗应用提供促进 EPC 扩增的合适表面。