Cell detachment is a routine process to recover seeded cells from culture systems. However, traditional enzymatic methods might easily induce senescence or apoptosis of cells during the routine passage process due to an improper handing process. The pH-responsive characteristic of chitosan has been demonstrated to control cell attachment/detachment by precise adjustment of medium pH within 1 h without additional enzymatic treatment and extensive washing steps. But the time 1 h is not satisfying to all applications to recover adherent cells from chitosan surface. The purpose of this study is to regulate pH-responsive characteristics of chitosan by blending with non-pH-responsive biomaterials, polycaprolactone (PCL) and polyvinyl alcohol (PVA) to provide a gentle rapid method for cell detachment. In this study, PCL could delay the pH-responsive characteristics of chitosan to reduce cell detachment ratio (< 65%) after raising medium pH to 7.65 for 2 h. In contrast, PVA could provide a synergistic effect on rapid detachment of cells (>95%) from the chitosan/PVA blended surface within 30 min after raising medium pH to 7.65. We further discuss the related mechanism among cell detachment, fibronectin, and chitosan-based blends. These results indicate non-pH-responsive PCL and PVA could be added into pH-responsive chitosan to develop a rapid non-enzymatic strategy to control cell detachment rate for cell passage and various tissue engineering applications.

细胞分离是从培养系统中回收接种细胞的常规过程。然而，由于不正确的处理过程，传统的酶促方法可能在常规传代过程中容易诱导细胞衰老或凋亡。壳聚糖的pH响应特性已证明可通过在1小时内精确调节培养基pH值来控制细胞的附着/脱离，而无需额外的酶处理和大量洗涤步骤。但是1小时的时间并不能满足所有从壳聚糖表面回收粘附细胞的应用。这项研究的目的是通过与非pH响应生物材料，聚己内酯（PCL）和聚乙烯醇（PVA）混合来调节壳聚糖的pH响应特性，以提供一种温和，快速的细胞分离方法。在这个研究中，将培养基的pH升高至7.65 2 h后，PCL可以延迟壳聚糖的pH响应特性，从而降低细胞脱离率（<65％）。相比之下，在将培养基的pH值提高到7.65后的30分钟内，PVA可以对细胞从壳聚糖/ PVA混合表面的快速脱离（> 95％）提供协同作用。我们进一步讨论了细胞脱离，纤连蛋白和壳聚糖基混合物之间的相关机制。这些结果表明，可以将非pH响应的PCL和PVA添加到pH响应的壳聚糖中，从而开发出一种快速的非酶促策略来控制细胞的脱落速率，以用于细胞传代和各种组织工程应用。在将中等pH值提高至7.65后的30分钟内，从壳聚糖/ PVA混合表面中提取95％的糖）。我们进一步讨论了细胞脱离，纤连蛋白和壳聚糖基混合物之间的相关机制。这些结果表明可以将非pH响应的PCL和PVA添加到pH响应的壳聚糖中，从而开发出一种快速的非酶促策略来控制细胞的脱落速率，以用于细胞传代和各种组织工程应用。在将培养基的pH值提高到7.65后的30分钟内，从壳聚糖/ PVA混合表面中提取95％的糖）。我们进一步讨论了细胞脱离，纤连蛋白和壳聚糖基混合物之间的相关机制。这些结果表明，可以将非pH响应的PCL和PVA添加到pH响应的壳聚糖中，从而开发出一种快速的非酶促策略来控制细胞的脱落速率，以用于细胞传代和各种组织工程应用。