Thermoresponsive Worms for Expansion and Release of Human Embryonic Stem Cells

• Published in: Biomacromolecules Vol. 15; no. 3; pp. 844 - 855
• Main Authors: Chen, Xiaoli, Prowse, Andrew B. J, Jia, Zhongfan, Tellier, Helena, Munro, Trent P, Gray, Peter P, Monteiro, Michael J
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 10.03.2014
• Subjects: Animal cells; Applied sciences; Biological and medical sciences; Biotechnology; Cell Culture Techniques; Cell Differentiation - genetics; Cell Proliferation; Embryoid Bodies - chemistry; Embryoid Bodies - cytology; Embryonic Stem Cells - cytology; Establishment of new cell lines, improvement of cultural methods, mass cultures; Eukaryotic cell cultures; Exact sciences and technology; Extracellular Matrix - chemistry; Fundamental and applied biological sciences. Psychology; Humans; Methods. Procedures. Technologies; Organic polymers; Physicochemistry of polymers; Polymers - chemistry; Polymers with particular properties; Preparation, kinetics, thermodynamics, mechanism and catalysts; Regenerative Medicine; Temperature; Cell proliferation; Biological properties; End group; Stem cell; Temperature effect; Cytotoxicity; Vitronectin; Acrylamide derivative copolymer; Fibronectin; Suspension culture; Chain transfer; Emulsion polymerization; Cell adhesion; Recombinant protein; Organic trithiocarbonate; Liquid solid adsorption; Embryoid body; Cell differentiation; Acrylamide derivative polymer; Biological activity; Monodispersed polymer; Free radical polymerization; Three dimensional culture; Embryonic cell; Preparation; Diblock copolymer; Nanostructured materials; Styrene copolymer
• DOI: 10.1021/bm401702h
• ISSN: 1525-7797

The development of robust suspension cultures of human embryonic stem cells (hESCs) without the use of cell membrane disrupting enzymes or inhibitors is critical for future clinical applications in regenerative medicine. We have achieved this by using long, flexible, and thermoresponsive polymer worms decorated with a recombinant vitronectin subdomain that bridge hESCs, aiding in hESC’s natural ability to form embryoid bodies (EBs) and satisfying their inherent requirement for cell–cell and cell–extracellular matrix contact. When the EBs reached an optimal upper size where cytokine and nutrient penetration becomes limiting, these long and flexible polymer worms facilitated EB breakdown via a temperature shift from 37 to 25 °C. The thermoresponsive nature of the worms enabled a cyclical dissociation and propagation of the cells. Repeating the process for three cycles (over eighteen days) provided a >30-fold expansion in cell number while maintaining pluripotency, thereby providing a simple, nondestructive process for the 3D expansion of hESC.