Three-dimensional porous alginate scaffolds provide a conducive environment for generation of well-vascularized embryoid bodies from human embryonic stem cells.
Differentiation of human embryonic stem cells (hESCs) can be instigated through the formation of embryo-like aggregates in suspension, termed human embryoid bodies (hEBs). Controlling cell aggregation and agglomeration during hEBs formation has a profound effect on the extent of cell proliferation and differentiation. In a previous work, we showed that control over hEBs formation and differentiation can be achieved via cultivation of hESC suspensions in a rotating bioreactor system. We now report that hEBs can be generated directly from hESC suspensions within three-dimensional (3D) porous alginate scaffolds. The confining environments of the alginate scaffold pores enabled efficient formation of hEBs with a relatively high degree of cell proliferation and differentiation; encouraged round, small-sized hEBs; and induced vasculogenesis in the forming hEBs to a greater extent than in static or rotating cultures. We therefore conclude that differentiation of hEBs can be induced and directed by physical constraints in addition to chemical cues.
Gerecht-Nir
Sharon
S
Biotechnology Interdisciplinary Unit, Technion-Israel Institute of Technology, Haifa, Israel.
Cohen
Smadar
S
Ziskind
Anna
A
Itskovitz-Eldor
Joseph
J
D000464:Alginates / Q000737:chemistry*; D021522:Bioartificial Organs; D001672:Biocompatible Materials / Q000737:chemistry; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D048429:Cell Size; D002470:Cell Survival / Q000502:physiology; D004622:Embryo, Mammalian / Q000098:blood supply* / Q000166:cytology*; D047108:Embryonic Development / Q000502:physiology*; D020723:Glucuronic Acid / Q000737:chemistry*; D006603:Hexuronic Acids / Q000737:chemistry*; D006801:Humans; D008422:Materials Testing; D008567:Membranes, Artificial; D018919:Neovascularization, Physiologic / Q000502:physiology; D016062:Porosity; D013234:Stem Cells / Q000166:cytology* / Q000502:physiology*; D023822:Tissue Engineering / Q000379:methods*
D000464:Alginates; D001672:Biocompatible Materials; D006603:Hexuronic Acids; D008567:Membranes, Artificial; D020723:Glucuronic Acid