title: Thermoresponsive worms for expansion and release of human embryonic stem cells.
abstract: 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.
authors.affiliation: Australian Institute for Bioengineering and Nanotechnology, The University of Queensland , Brisbane QLD 4072, Australia.
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Chen
authors.forename: Andrew B J
authors.initials: AB
authors.lastname: Prowse
authors.forename: Zhongfan
authors.initials: Z
authors.lastname: Jia
authors.forename: Helena
authors.initials: H
authors.lastname: Tellier
authors.forename: Trent P
authors.initials: TP
authors.lastname: Munro
authors.forename: Peter P
authors.initials: PP
authors.lastname: Gray
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Monteiro
mesh_terms: D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000235:genetics; D049109:Cell Proliferation; D058732:Embryoid Bodies / Q000737:chemistry* / Q000166:cytology; D053595:Embryonic Stem Cells / Q000166:cytology*; D005109:Extracellular Matrix / Q000737:chemistry*; D006801:Humans; D011108:Polymers / Q000737:chemistry; D044968:Regenerative Medicine; D013696:Temperature
chemical_list: D011108:Polymers