Kasai-Brunswick Taís Hanae, Costa Andréa Rodrigues da, Barbosa Raiana Andrade Quintanilha, Farjun Bruna, Mesquita Fernanda Cristina Paccola, Silva Dos Santos Danúbia, Ramos Isalira Peroba, Suhett Grazielle, Brasil Guilherme Visconde, Cunha Sandro Torrentes da, Brito José Oscar R, Passipieri Juliana do Amaral, Carvalho Adriana Bastos, Campos de Carvalho Antonio Carlos
Institute of Biophysics Carlos Chagas Filho, Federal University of Rio de Janeiro, Av. Carlos Chagas Filho, n°373, room G2-053, CEP:21941-902, Rio de Janeiro, RJ, Brazil.
National Institute of Cardiology, Rua das Laranjeiras, n°374-Laranjeiras, CEP:22240-006, Rio de Janeiro, RJ, Brazil.
Stem Cell Res Ther. 2017 Feb 15;8(1):36. doi: 10.1186/s13287-017-0481-x.
Heart failure represents an important public health issue due to its high costs and growing incidence worldwide. Evidence showing the regenerative potential of postmitotic heart tissue has suggested the existence of endogenous cardiac stem cells in adult hearts. Cardiosphere-derived cells (CDC) constitute a candidate pool of such cardiac stem cells. Previous studies using acute myocardial infarction (MI) models in rodents demonstrated an improvement in cardiac function after cell therapy with CDC. We evaluated the therapeutic potential of CDC 60 days after MI in a rat model.
CDC were obtained from human discarded myocardial tissue and rat hearts by enzymatic digestion with collagenase II. At 10-15 days after isolation, small, round, phase-bright cells (PBCs) appeared on top of the adherent fibroblast-like cells. The PBCs were collected and placed on a nonadherent plate for 2 days, where they formed cardiospheres which were then transferred to adherent plates, giving rise to CDC. These CDC were characterized by flow cytometry. Wistar rats were submitted to MI through permanent occlusion of the anterior descending coronary artery. After 60 days, they were immunosuppressed with cyclosporine A during 10 days. On the third day, infarcted animals were treated with 5 × 10 human CDC (hCDC) or placebo through intramyocardial injection guided by echocardiogram. Another group of animals was treated with rat CDC (rCDC) without immunosuppression. hCDC and rCDC were stably transduced with a viral construct expressing luciferase under control of a constitutive promoter. CDC were then used in a bioluminescence assay. Functional parameters were evaluated by echocardiogram 90 and 120 days after MI and by Langendorff at 120 days.
CDC had a predominantly mesenchymal phenotype. Cell tracking by bioluminescence demonstrated over 85% decrease in signal at 5-7 days after cell therapy. Cardiac function evaluation by echocardiography showed no differences in ejection fraction, end-diastolic volume, or end-systolic volume between groups receiving human cells, rat cells, or placebo. Hemodynamic analyses and infarct area quantification confirmed that there was no improvement in cardiac remodeling after cell therapy with CDC.
Our study challenges the effectiveness of CDC in post-ischemic heart failure.
心力衰竭因其高昂的成本和在全球范围内不断上升的发病率，成为一个重要的公共卫生问题。有证据表明有丝分裂后心脏组织具有再生潜力，这提示成年心脏中存在内源性心脏干细胞。心脏球衍生细胞（CDC）构成了此类心脏干细胞的一个候选库。先前使用啮齿动物急性心肌梗死（MI）模型的研究表明，用CDC进行细胞治疗后心脏功能有所改善。我们在大鼠模型中评估了心肌梗死后60天CDC的治疗潜力。
通过用胶原酶II进行酶消化，从人类废弃心肌组织和大鼠心脏中获取CDC。在分离后10 - 15天，小的、圆形的、相位明亮的细胞（PBCs）出现在贴壁的成纤维细胞样细胞之上。收集PBCs并置于非贴壁培养板上2天，在此期间它们形成心脏球，然后将其转移至贴壁培养板，从而产生CDC。这些CDC通过流式细胞术进行表征。通过永久性结扎前降支冠状动脉使Wistar大鼠发生心肌梗死。60天后，它们用环孢素A免疫抑制10天。在第3天，通过超声心动图引导，对梗死动物进行心肌内注射5×10个人类CDC（hCDC）或安慰剂治疗。另一组动物用大鼠CDC（rCDC）治疗，不进行免疫抑制。hCDC和rCDC用在组成型启动子控制下表达荧光素酶的病毒构建体进行稳定转导。然后将CDC用于生物发光测定。在心肌梗死后90天和120天通过超声心动图以及在120天通过Langendorff法评估功能参数。
CDC主要具有间充质表型。通过生物发光进行细胞追踪显示，细胞治疗后5 - 7天信号下降超过85%。通过超声心动图评估心脏功能显示，接受人类细胞、大鼠细胞或安慰剂的组之间在射血分数、舒张末期容积或收缩末期容积方面没有差异。血流动力学分析和梗死面积定量证实，用CDC进行细胞治疗后心脏重塑没有改善。
我们的研究对CDC在缺血性心力衰竭后的有效性提出了质疑。
