Kim Yu-Jin, Lee Tae-Jin, Jeong Gun-Jae, Song Jihun, Yu Taekyung, Lee Doo Sung, Bhang Suk Ho
School of Chemical Engineering, Sungkyunkwan University, Suwon 16419, Korea.
Department of Bio-Health Convergence, Kangwon National University, Chuncheon 24341, Korea.
Materials (Basel). 2021 Jan 20;14(3):491. doi: 10.3390/ma14030491.
Cell therapy usually accompanies cell detachment as an essential process in cell culture and cell collection for transplantation. However, conventional methods based on enzymatic cell detachment can cause cellular damage including cell death and senescence during the routine cell detaching step due to an inappropriate handing. The aim of the current study is to apply the pH-responsive degradation property of poly (amino ester) to the surface of a cell culture dish to provide a simple and easy alternative method for cell detachment that can substitute the conventional enzyme treatment. In this study, poly (amino ester) was modified (cell detachable polymer, CDP) to show appropriate pH-responsive degradation under mild acidic conditions (0.05% (w/v) CDP, pH 6.0) to detach stem cells (human adipose tissue-derived stem cells (hADSCs)) perfectly within a short period (less than 10 min). Compared to conventional enzymatic cell detachment, hADSCs cultured on and detached from a CDP-coated cell culture dish showed similar cellular properties. We further performed in vivo experiments on a mouse hindlimb ischemia model (1.0 × 10 cells per limb). The in vivo results indicated that hADSCs retrieved from normal cell culture dishes and CDP-coated cell culture dishes showed analogous therapeutic angiogenesis. In conclusion, CDP could be applied to a pH-responsive cell detachment system as a simple and easy nonenzymatic method for stem cell culture and various cell therapies.
细胞治疗通常伴随着细胞脱离，这是细胞培养和用于移植的细胞收集过程中的一个基本步骤。然而，基于酶促细胞脱离的传统方法在常规细胞脱离步骤中，由于操作不当，可能会导致包括细胞死亡和衰老在内的细胞损伤。本研究的目的是将聚（氨基酯）的pH响应降解特性应用于细胞培养皿表面，以提供一种简单易行的替代方法来进行细胞脱离，从而取代传统的酶处理方法。在本研究中，聚（氨基酯）被修饰（细胞可脱离聚合物，CDP），使其在温和酸性条件（0.05%（w/v）CDP，pH 6.0）下表现出适当的pH响应降解，从而在短时间内（不到10分钟）完美地脱离干细胞（人脂肪组织来源的干细胞（hADSCs））。与传统的酶促细胞脱离相比，在CDP包被的细胞培养皿上培养并从其上脱离的hADSCs表现出相似的细胞特性。我们进一步在小鼠后肢缺血模型（每只肢体1.0×10个细胞）上进行了体内实验。体内实验结果表明，从正常细胞培养皿和CDP包被的细胞培养皿中获取的hADSCs表现出类似的治疗性血管生成作用。总之，CDP可作为一种简单易行的非酶方法应用于pH响应性细胞脱离系统，用于干细胞培养和各种细胞治疗。
