Purified cardiomyocytes from bone marrow mesenchymal stem cells produce stable intracardiac grafts in mice
Purified cardiomyocytes from bone marrow mesenchymal stem cells produce stable intracardiac grafts in mice
复制标题
DOI：
10.1016/j.cardiores.2004.10.004
复制
发表时间：
2005-02-01
影响因子：
10.8
通讯作者：
Fukuda, K
中科院分区：
文献类型：
--
作者：
Hattan, N;Kawaguchi, H;Fukuda, K
目的：我们曾从小鼠骨髓中分离出心肌细胞(CMG细胞)。再生的心肌细胞是细胞移植的重要候选细胞，但由于它们是干细胞来源的，它们可能会被各种类型的细胞污染，因此需要鉴定和纯化。我们的目的是提高细胞移植的效率，保护受体免受可能的不良反应的影响，并对再生的心肌细胞进行鉴定。方法：在肌球蛋白轻链-2v(MLC-2v)启动子的控制下，使用荧光激活的细胞分选器从干细胞来源的混合物中分离非心肌细胞，特异性地分离携带增强型绿色荧光蛋白(EGFP)基因的重组质粒的CMG细胞。结果：诱导分化后6%~24%的转基因CMG细胞表达EGFP，并筛选出较强的EGFP阳性表达组分。所有分离的细胞在3周后开始自发搏动。这些细胞表达心肌细胞特异性基因，如α-骨骼肌动蛋白、β-肌球蛋白重链、MLC-2v和CaV1.2，并掺入溴脱氧尿苷5天。将分离的EGFP阳性细胞扩增5d后移植到成年小鼠的左心室内。移植细胞存活至少3个月，并与受体心脏的心肌细胞平行定向。结论：从成体干细胞中纯化和移植分化的心肌细胞为治疗心力衰竭提供了一种可行的组织工程模型。(C)2004年欧洲心脏病学会。爱思唯尔出版公司版权所有。
Objective: We have previously isolated cardiomyogenic cells from murine bone marrow (CMG cells). Regenerated cardiomyocytes are important candidates for cell transplantation, but as they are stem cell derived, they can be contaminated with various cell types, thereby requiring characterization and purification. Our objectives were to increase the efficiency of cell transplantation and to protect the recipients from possible adverse effects using an efficient and effective purification process as well as to characterize regenerated cardiomyocytes.Methods: Noncardiomyocytes were eliminated from a mixture of stem-cell-derived cells using a fluorescence-activated cell sorter to specifically isolate CMG cells transfected with a recombinant plasmid containing enhanced green fluorescent protein (EGFP) cDNA under the control of the myosin light chain-2v (MLC-2v) promoter. Gene expression and the action potential were investigated, and purified cells were transplanted into the heart of adult mice.Results: Six percent to 24% of transfected CMG cells expressed EGFP after differentiation was induced, and a strong EGFP-positive fraction was selected. All the sorted cells began spontaneous beating after 3 weeks. These cells expressed cardiomyocyte-specific genes such as alpha-skeletal actin, beta-myosin heavy chain, MLC-2v, and CaV1.2 and incorporated bromodeoxyuridine for 5 days. The isolated EGFP-positive cells were expanded for 5 days and then transplanted into the left ventricle of adult mouse hearts. The transplanted cells survived for at least 3 months and were oriented in parallel to the cardiomyocytes of the recipient heart.Conclusions: The purification and transplantation of differentiated cardiomyocytes from adult stem cells provides a viable model of tissue engineering for the treatment of heart failure. (C) 2004 European Society of Cardiology. Published by Elsevier B.V All rights reserved.