Choi Goro, Cho Younghak, Yu Seung Jung, Baek Jieung, Lee Minseok, Kim Yesol, Lee Eunjung, Im Sung Gap
Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea.
ACS Appl Bio Mater. 2020 Nov 16;3(11):7654-7665. doi: 10.1021/acsabm.0c00844. Epub 2020 Oct 13.
For efficient therapeutic use of human mesenchymal stem cells (hMSCs), maximizing their self-renewal performance and multipotency must be fully retained. However, conventional trypsin-based cell passaging methods are known to damage the attached cells to be detached because of the inherent corrosive nature of trypsin, and continuous passaging substantially degrades the self-renewal and differentiation capacity of hMSCs. Therefore, it is imperative to secure a damage-free passaging method that supports cell growth as well as their stem cell function. Here, an enzyme-free cell detachment method using a poly(ethylene glycol dimethacrylate) (pEGDMA)-coated surface is developed, which allows for reduced integrin-dependent cell adhesion. Cell detachment can be facilitated simply by treating the plated cells on the pEGDMA surface with Ca and Mg-depleted DPBS. Spontaneous cell detachment occurs within 10 min with the full retention of the cell viability and proliferation ability of hMSCs. Especially, the detachment method can minimize the surface protein damage of hMSCs compared to the conventional trypsin treatment and preserve the self-renewal property and differentiation capacity even with an increased passage number over 10. The developed enzyme-free detachment method using the pEGDMA-coated surface is highly promising for a culture platform to broaden its application to the field of tissue engineering and regenerative medicine.
为了高效治疗性应用人间充质干细胞（hMSCs），必须充分保留其自我更新能力和多能性。然而，已知传统的基于胰蛋白酶的细胞传代方法会因胰蛋白酶固有的腐蚀性而损伤待分离的贴壁细胞，连续传代还会显著降低hMSCs的自我更新和分化能力。因此，必须找到一种无损传代方法，既能支持细胞生长，又能维持其干细胞功能。在此，开发了一种使用聚（乙二醇二甲基丙烯酸酯）（pEGDMA）涂层表面的无酶细胞分离方法，该方法可减少整合素依赖性细胞黏附。只需用不含钙和镁的DPBS处理pEGDMA表面上接种的细胞，就能促进细胞分离。hMSCs的细胞活力和增殖能力得以完全保留，细胞在10分钟内即可自发分离。特别是，与传统的胰蛋白酶处理相比，这种分离方法能将hMSCs的表面蛋白损伤降至最低，即使传代次数超过10次，仍能保持自我更新特性和分化能力。利用pEGDMA涂层表面开发的这种无酶分离方法，对于拓宽其在组织工程和再生医学领域应用的培养平台极具前景。
