TY - JOUR
T1 - Chitosan 3D cell culture system promotes naïve-like features of human induced pluripotent stem cells
T2 - A novel tool to sustain pluripotency and facilitate differentiation
AU - Chang, Po Hsiang
AU - Chao, Hsiao Mei
AU - Chern, Edward
AU - Hsu, Shan hui
N1 - Funding Information:
This study was supported by grants from the Program of Regenerative Medicine, Ministry of Science and Technology (MOST), Taiwan (MOST 109-2321-B-002-044, MOST 109-2320-B-002-051-MY2 and NSC 100-2314-B-002-002). We are grateful for Dr. Su-Yi Tsai to assist the procedure of cardiomyocyte differentiation. We also thank the excellent technical support of Technology Commons, College of Life Science, National Taiwan University.
Funding Information:
This study was supported by grants from the Program of Regenerative Medicine, Ministry of Science and Technology ( MOST ), Taiwan ( MOST 109-2321-B-002-044 , MOST 109-2320-B-002-051-MY2 and NSC 100-2314-B-002-002 ). We are grateful for Dr. Su-Yi Tsai to assist the procedure of cardiomyocyte differentiation. We also thank the excellent technical support of Technology Commons, College of Life Science, National Taiwan University.
Publisher Copyright:
© 2020 Elsevier Ltd
PY - 2021/1
Y1 - 2021/1
N2 - 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.
AB - 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.
KW - 3D feeder-free culture system
KW - Chitosan
KW - Human induced pluripotent stem cell
KW - Naïve pluripotency
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U2 - 10.1016/j.biomaterials.2020.120575
DO - 10.1016/j.biomaterials.2020.120575
M3 - Article
C2 - 33341735
AN - SCOPUS:85097713826
SN - 0142-9612
VL - 268
JO - Biomaterials
JF - Biomaterials
M1 - 120575
ER -