Wang J S, Shum-Tim D, Galipeau J, Chedrawy E, Eliopoulos N, Chiu R C
Division of Cardiothoracic Surgery, McGill University, the Division of Hematology Oncology, Jewish General Hospital/McGill University, Montreal, Quebec, Canada.
J Thorac Cardiovasc Surg. 2000 Nov;120(5):999-1005. doi: 10.1067/mtc.2000.110250.
Marrow stromal cells are mesenchymal stem cells able to differentiate into cardiomyocytes in vitro. We tested the hypothesis that marrow stromal cells, when implanted into myocardium, can undergo milieu-dependent differentiation and express cardiomyogenic phenotypes in vivo.
Isogenic adult rats were used as donors and recipients to simulate autologous transplantation. Marrow stromal cells isolated from donor leg bones were culture-expanded, labeled with 4;,6-diamidino-2-phenylindole, and then injected into the myocardium of the recipients. The hearts were harvested from 4 days to 12 weeks after implantation, and the implant sites were examined to identify the phenotypes of the labeled marrow stromal cells.
Viable cells labeled with 4;, 6-diamidino-2-phenylindole can be identified in host myocardium at all time points after implantation. Implanted marrow stromal cells show the growth potential in a myocardial environment. After 4 weeks, donor cells derived from marrow stromal cells demonstrate myogenic differentiation with the expression of sarcomeric myosin heavy chain and organized contractile proteins. Positive staining for connexin 43 indicates the formation of gap junctions, which suggests that cells derived from marrow stromal cells, as well as native cardiomyocytes, are connected by intercalated disks.
Different cell sources have been used as donor cells for cellular cardiomyoplasty. Our findings indicate that marrow stromal cells can also be used as donor cells. In an appropriate microenvironment they will exhibit cardiomyogenic phenotypes and may replace native cardiomyocytes lost by necrosis or apoptosis. Because marrow stromal cells can be obtained repeatedly by bone marrow aspiration and expanded vastly in vitro before being implanted or used as autologous implants, and because their use does not call for immunosuppression, the clinical use of marrow stromal cells for cellular cardiomyoplasty appears to be most advantageous.
骨髓基质细胞是能够在体外分化为心肌细胞的间充质干细胞。我们检验了这样一个假说,即骨髓基质细胞植入心肌后,能够在体内经历环境依赖性分化并表达心肌生成表型。
采用同基因成年大鼠作为供体和受体以模拟自体移植。从供体腿骨分离出的骨髓基质细胞经培养扩增,用4′,6-二脒基-2-苯基吲哚标记,然后注入受体的心肌内。在植入后4天至12周采集心脏,检查植入部位以确定标记的骨髓基质细胞的表型。
在植入后的所有时间点,均可在宿主心肌中鉴定出用4′,6-二脒基-2-苯基吲哚标记的活细胞。植入的骨髓基质细胞在心肌环境中显示出生长潜力。4周后,源自骨髓基质细胞的供体细胞表现出肌源性分化,伴有肌节肌球蛋白重链和有组织的收缩蛋白的表达。连接蛋白43的阳性染色表明形成了缝隙连接,这提示源自骨髓基质细胞的细胞以及天然心肌细胞通过闰盘相连。
不同的细胞来源已被用作细胞心肌成形术的供体细胞。我们的研究结果表明,骨髓基质细胞也可作为供体细胞。在合适的微环境中,它们将表现出心肌生成表型,并可能替代因坏死或凋亡而丢失的天然心肌细胞。由于骨髓基质细胞可通过骨髓穿刺反复获取,并在植入前或用作自体植入物之前在体外大量扩增,且使用它们不需要免疫抑制,因此骨髓基质细胞用于细胞心肌成形术的临床应用似乎最为有利。