人脐带间充质干细胞片治疗缺血性心脏组织的临床前研究。

BACKGROUND: Human umbilical cord mesenchymal stem cells (hUC-MSCs) have been widely used due to their multipotency, a broad range of sources, painless collection, and compliance with standard amplification. Cell sheet technology is a tissue engineering methodology requiring scaffolds free, and it provides an effective method for cell transplantation. To improve the therapeutic efficacy, we combined hUC-MSCs with cell sheet technology to evaluate the safety and efficacy of hUC-MSC sheets in preclinical studies using appropriate animal models. METHODS: hUC-MSC sheets were fabricated by hUC-MSCs from a cell bank established by a standard operation process and quality control. Cytokine secretion, immunoregulation, and angiopoiesis were evaluated in vitro. Oncogenicity and cell diffusion assays of hUC-MSC sheets were conducted to verify the safety of hUC-MSCs sheet transplantation in mice. To provide more meaningful animal experimental data for clinical trials, porcine myocardial infarction (MI) models were established by constriction of the left circumflex, and hUC-MSC sheets were transplanted onto the ischemic area of the heart tissue. Cardiac function was evaluated and compared between the experimental and MI groups. RESULTS: The in vitro results showed that hUC-MSC sheets could secrete multiple cellular factors, including VEGF, HGF, IL-6, and IL-8. Peripheral blood mononuclear cells had a lower proliferation rate and lower TNF-α secretion when co-cultured with hUC-MSC sheets. TH1 cells had a smaller proportion after activation. In vivo safety results showed that the hUC-MSCs sheet had no oncogenicity and was mainly distributed on the surface of the ischemic myocardial tissue. Echocardiography showed that hUC-MSC sheets effectively improved the left ventricular ejection fraction (LVEF), and the LVEF significantly changed (42.25 ± 1.23% vs. 66.9 ± 1.10%) in the hUC-MSC transplantation group compared with the MI group (42.52 ± 0.65% vs. 39.55 ± 1.97%) at 9 weeks. The infarct ratio of the hUC-MSCs sheet transplantation groups was also significantly reduced (14.2 ± 4.53% vs. 4.00 ± 2.00%) compared with that of the MI group. CONCLUSION: Allogeneic source and cell bank established by the standard operation process and quality control make hUC-MSCs sheet possible to treat MI by off-the-shelf drug with universal quality instead of individualized medical technology.

背景：人脐带间充质干细胞（hUC-MSCs）因其多能性、广泛的来源、无痛采集和符合标准扩增而被广泛应用。细胞片技术是一种组织工程方法，需要无支架，为细胞移植提供了一种有效的方法。为了提高治疗效果，我们将 hUC-MSCs 与细胞片技术相结合，使用适当的动物模型，在临床前研究中评估 hUC-MSC 片的安全性和有效性。

方法：hUC-MSC 片由通过标准操作流程和质量控制建立的细胞库中的 hUC-MSCs 制成。体外评估细胞因子分泌、免疫调节和血管生成。进行 hUC-MSC 片的致癌性和细胞扩散试验，以验证 hUC-MSC 片在小鼠中的移植安全性。为了为临床试验提供更有意义的动物实验数据，通过左回旋支结扎建立猪心肌梗死（MI）模型，并将 hUC-MSC 片移植到心脏组织的缺血区域。比较实验组和 MI 组之间的心脏功能。

结果：体外结果表明，hUC-MSC 片可分泌多种细胞因子，包括 VEGF、HGF、IL-6 和 IL-8。与 hUC-MSC 片共培养时，外周血单核细胞的增殖率较低，TNF-α分泌较低。TH1 细胞激活后比例较小。体内安全性结果表明，hUC-MSCs 片无致癌性，主要分布在缺血心肌组织表面。超声心动图显示，hUC-MSC 片有效提高了左心室射血分数（LVEF），与 MI 组（42.52±0.65%比 39.55±1.97%）相比，hUC-MSC 移植组的 LVEF 明显变化（42.25±1.23%比 66.9±1.10%）在 9 周时。hUC-MSCs 片移植组的梗死比例也明显低于 MI 组（14.2±4.53%比 4.00±2.00%）。

结论：同种异体来源和通过标准操作流程和质量控制建立的细胞库使 hUC-MSCs 片有可能通过现成的药物治疗 MI，而不是个体化的医疗技术。