A simplified and cost-effective culture system for maintaining the pluripotency of human induced pluripotent stem cells (hiPSCs) is crucial for stem cell applications. Although recombinant protein-based feeder-free hiPSC culture systems have been developed, their manufacturing processes are expensive and complicated, which hinders hiPSC technology progress. Chitosan, a versatile biocompatible polysaccharide, has been reported as a biomaterial for three-dimensional (3D) cell culture system that promotes the physiological activities of mesenchymal stem cells and cancer cells. In the current study, we demonstrated that chitosan membranes sustained proliferation and pluripotency of hiPSCs in long-term culture (up to 365 days). Moreover, using vitronectin as the comparison group, the pluripotency of hiPSCs grown on the membranes was altered into a naïve-like state, which, for pluripotent stem cells, is an earlier developmental stage with higher stemness. On the chitosan membranes, hiPSCs self-assembled into 3D spheroids with an average diameter of ~100 μm. These hiPSC spheroids could be directly differentiated into lineage-specific cells from the three germ layers with 3D structures. Collectively, chitosan membranes not only promoted the naïve pluripotent features of hiPSCs but also provided a novel 3D differentiation platform. This convenient biomaterial-based culture system may enable the effective expansion and accessibility of hiPSCs for regenerative medicine, disease modeling, and drug screening.

维持人类诱导的多能干细胞（hiPSC）的多能性的简化且具有成本效益的培养系统对于干细胞应用至关重要。尽管已经开发了基于重组蛋白的无饲养层的hiPSC培养系统，但是它们的制造过程昂贵且复杂，这阻碍了hiPSC技术的发展。壳聚糖是一种通用的生物相容性多糖，据报道是三维（3D）细胞培养系统的生物材料，可促进间充质干细胞和癌细胞的生理活性。在当前的研究中，我们证明了壳聚糖膜在长期培养（长达365天）中能够维持hiPSC的增殖和多能性。此外，以玻连蛋白为对照组，在膜上生长的hiPSC的多能性被改变为幼稚的状态，对于多能干细胞而言，它是具有更高干性的早期发育阶段。在壳聚糖膜上，hiPSCs自组装成平均直径约为100μm的3D球体。这些hiPSC球状体可以从具有3D结构的三个胚层直接分化为谱系特异性细胞。总的来说，壳聚糖膜不仅促进了hiPSC的幼稚多能性特征，而且还提供了一个新颖的3D分化平台。这种方便的基于生物材料的培养系统可以实现hiPSC的有效扩展和可访问性，用于再生医学，疾病建模和药物筛选。在壳聚糖膜上，hiPSCs自组装成平均直径约为100μm的3D球体。这些hiPSC球状体可以从具有3D结构的三个胚层直接分化为谱系特异性细胞。总的来说，壳聚糖膜不仅促进了hiPSC的幼稚多能性特征，而且还提供了一个新颖的3D分化平台。这种方便的基于生物材料的培养系统可以实现hiPSC的有效扩展和可访问性，用于再生医学，疾病建模和药物筛选。在壳聚糖膜上，hiPSCs自组装成平均直径约为100μm的3D球体。这些hiPSC球状体可以从具有3D结构的三个胚层直接分化为谱系特异性细胞。总的来说，壳聚糖膜不仅促进了hiPSC的幼稚多能性特征，而且还提供了一个新颖的3D分化平台。这种方便的基于生物材料的培养系统可以实现hiPSC的有效扩展和可访问性，用于再生医学，疾病建模和药物筛选。壳聚糖膜不仅促进了hiPSC的幼稚多能性，而且提供了一个新颖的3D分化平台。这种方便的基于生物材料的培养系统可以实现hiPSC的有效扩展和可访问性，用于再生医学，疾病建模和药物筛选。壳聚糖膜不仅促进了hiPSC的幼稚多能性，而且提供了一个新颖的3D分化平台。这种方便的基于生物材料的培养系统可以实现hiPSC的有效扩展和可访问性，用于再生医学，疾病建模和药物筛选。