Binary colloidal crystals (BCCs) as a feeder-free system to generate human induced pluripotent stem cells (hiPSCs).
Human induced pluripotent stem cells (hiPSCs) are capable of differentiating into any cell type and provide significant advances to cell therapy and regenerative medicine. However, the current protocol for hiPSC generation is relatively inefficient and often results in many partially reprogrammed colonies, which increases the cost and reduces the applicability of hiPSCs. Biophysical stimulation, in particular from tuning cell-surface interactions, can trigger specific cellular responses that could in turn promote the reprogramming process. In this study, human fibroblasts were reprogrammed into hiPSCs using a feeder-free system and episomal vectors using novel substrates based on binary colloidal crystals (BCCs). BCCs are made from two different spherical particle materials (Si and PMMA) ranging in size from nanometers to micrometers that self-assemble into hexagonal close-packed arrays. Our results show that the BCCs, particularly those made from a crystal of 2 μm Si and 0.11 μm PMMA particles (2SiPM) facilitate the reprogramming process and increase the proportion of fully reprogrammed hiPSC colonies, even without a vitronectin coating. Subsequent isolation of clonal hiPSC lines demonstrates that they express pluripotent markers (OCT4 and TRA-1-60). This proof-of-concept study demonstrates that cell reprogramming can be improved on substrates where surface properties are tailored to the application.
Department of Chemistry and Biotechnology, Faculty of Science, Engineering and Technology, Swinburne University of Technology, Hawthorn, Victoria 3122, Australia.|Graduate Institute of Nanomedicine and Medical Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei 110, Taiwan.|Department of Anatomy and Neuroscience, Florey Neuroscience and Mental Health Institute, The University of Melbourne, Victoria 3000, Australia.|CSIRO Manufacturing, Bayview Avenue, Clayton, 3168 Victoria, Australia.
Peng-Yuan
PY
Wang
Centre for Eye Research Australia, Royal Victorian Eye and Ear Hospital; Ophthalmology, Department of Surgery, University of Melbourne, Victoria 3002, Australia.
Sandy Shen-Chi
SS
Hung
CSIRO Manufacturing, Bayview Avenue, Clayton, 3168 Victoria, Australia.
Helmut
H
Thissen
Department of Chemistry and Biotechnology, Faculty of Science, Engineering and Technology, Swinburne University of Technology, Hawthorn, Victoria 3122, Australia.
Peter
P
Kingshott
Centre for Eye Research Australia, Royal Victorian Eye and Ear Hospital; Ophthalmology, Department of Surgery, University of Melbourne, Victoria 3002, Australia.
Raymond Ching-Bong
RC
Wong
D018929:Cell Culture Techniques; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D003102:Colloids; D003460:Crystallization; D003470:Culture Media / Q000737:chemistry; D005347:Fibroblasts / Q000502:physiology*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000502:physiology*; D012822:Silicon Dioxide / Q000737:chemistry*
D003102:Colloids; D003470:Culture Media; D012822:Silicon Dioxide