To maintain the cellular functions of a stem cells for therapeutic tissue engineering, an advanced cell culture method for safe cell dissociation is necessary. We developed a novel cell dissociation method by applying pH-responsive bioreducible polymer on the surface of tissue culture plates (TCPs). We applied acid-responsive bioreducible poly (amino ester) (PAE) as a new candidate for surface coating method to develop alternative cell dissociation method against conventional enzyme (trypsin-ethylene diamine tetra acetic acid (EDTA)) treatment. Human adipose derived stem cells (hADSCs) were cultured on and dissociated from PAE-coated TCPs to compare cell adhesion, cell proliferation, cell viability, and functionality to those of the cells cultured on and dissociated with trypsin-EDTA from normal TCPs without PAE coating. To confirm the in vivo therapeutic efficacy of the hADSCs retrieved from PAE-coated TCPs compared to that of the cells retrieved from normal TCPs with trypsin-EDTA, we induced skin defects at the dorsal area of mice and injected the cells collected from both conditions. With the PAE coating method, cell adhesion, cell proliferation, cell viability, and functionality, especially the angiogenic efficacy, were well preserved when compared to those of the cells treated with trypsin-EDTA. In addition to in vitro results, injecting hADSCs retrieved from PAE-coated TCPs showed similar in vivo angiogenesis and wound closing efficiency compared to those of injecting hADSCs retrieved from normal TCPs with trypsin-EDTA treatment at 2 weeks after the transplantation into mouse skin wound models. We proposed the alternative method for the cell dissociation with pH-responsive bioreducible polymer, PAE. This PAE coating method may lead to the development of alternative cell dissociation method without using enzyme for future regenerative medicine and stem cell therapy.

为了维持干细胞用于治疗性组织工程的细胞功能，需要用于安全细胞解离的先进细胞培养方法。我们通过在组织培养板（TCP）的表面上应用pH响应性可生物还原的聚合物，开发了一种新型的细胞解离方法。我们应用酸响应性生物可还原聚（氨基酯）（PAE）作为表面涂层方法的新候选者，以开发针对常规酶（胰蛋白酶-乙二胺四乙酸（EDTA））处理的替代细胞解离方法。将人脂肪来源的干细胞（hADSCs）培养在涂有PAE的TCP上并从其上解离，以比较与未涂有PAE涂层的正常TCP上的胰蛋白酶-EDTA进行培养和与胰蛋白酶-EDTA分离的细胞的粘附，细胞增殖，细胞活力和功能。 。为了证实与用胰蛋白酶-EDTA从正常TCPs检索到的细胞相比，从PAE包被的TCPs检索到的hADSCs的体内治疗功效，我们在小鼠的背部区域诱导了皮肤缺损，并注射了两种条件下收集的细胞。与用胰蛋白酶-EDTA处理的细胞相比，使用PAE包被方法可以很好地保留细胞粘附，细胞增殖，细胞活力和功能性，尤其是血管生成功效。除体外结果外，与从正常TCP回收的hADSC注射胰蛋白酶-EDTA处理在移植到小鼠皮肤伤口模型后2周相比，从PAE包覆的TCP回收的hADSC注射显示出相似的体内血管生成和伤口闭合效率。我们提出了一种与pH响应的生物可还原聚合物PAE进行细胞解离的替代方法。这种PAE包被方法可能会导致替代细胞解离方法的发展，而无需在未来的再生医学和干细胞疗法中使用酶。