Department of Chemical and Materials Engineering, National Central University, No. 300, Jhongda RD., Jhongli, Taoyuan 32001, Taiwan, ROC.
Department of Chemical Engineering, R&D Center for Membrane Technology, Chung Yuan Christian University, 200, Chung-Bei Rd., Chungli, Taoyuan 320, Taiwan, ROC.
Biomaterials. 2016 Jan;76:76-86. doi: 10.1016/j.biomaterials.2015.10.039. Epub 2015 Oct 21.
Stem cell culture is typically based on batch-type culture, which is laborious and expensive. Here, we propose a continuous harvest method for stem cells cultured on thermoresponsive nanobrush surfaces. In this method, stem cells are partially detached from the nanobrush surface by reducing the temperature of the culture medium below the critical solution temperature needed for thermoresponse. The detached stem cells are harvested by exchange into fresh culture medium. Following this, the remaining cells are continuously cultured by expansion in fresh culture medium at 37 °C. Thermoresponsive nanobrush surfaces were prepared by coating block copolymers containing polystyrene (for hydrophobic anchoring onto culture dishes) with three types of polymers: (a) polyacrylic acid with cell-binding oligopeptides, (b) thermoresponsive poly-N-isopropylacrylamide, and (c) hydrophilic poly(ethyleneglycol)methacrylate. The optimal coating durations and compositions for these copolymers to facilitate adequate attachment and detachment of human adipose-derived stem cells (hADSCs) and embryonic stem cells (hESCs) were determined. hADSCs and hESCs were continuously harvested for 5 and 3 cycles, respectively, via the partial detachment of cells from thermoresponsive nanobrush surfaces.
干细胞培养通常基于批处理式培养，这种方法既繁琐又昂贵。在这里，我们提出了一种在热响应纳米刷表面上培养干细胞的连续收获方法。在这种方法中，通过将培养基的温度降低到热响应所需的临界溶液温度以下，使干细胞从纳米刷表面部分脱离。通过将其交换到新鲜的培养基中，可以收获脱离的干细胞。然后，通过在 37°C 的新鲜培养基中进行扩展来连续培养剩余的细胞。通过涂覆含有聚苯乙烯（用于疏水锚定到培养皿上）的嵌段共聚物来制备热响应纳米刷表面，这些嵌段共聚物具有三种聚合物：(a)带有细胞结合寡肽的聚丙烯酸、(b) 热响应性聚 N-异丙基丙烯酰胺和 (c) 亲水性聚（甲基丙烯酸乙二醇酯）。确定了这些共聚物的最佳涂覆时间和组成，以促进人脂肪来源干细胞（hADSCs）和胚胎干细胞（hESCs）的充分附着和脱离。通过从热响应纳米刷表面部分脱离细胞，hADSCs 和 hESCs 分别连续收获了 5 次和 3 次循环。
