title: The use of agarose microwells for scalable embryoid body formation and cardiac differentiation of human and murine pluripotent stem cells.
abstract: In most pluripotent stem cell differentiation protocols the formation of embryoid bodies (EBs) is an important step. Here we describe a rapid, straightforward soft lithography approach for the preparation of hydrophilic silicon masters from different templates and the subsequent production of patterned agarose-DMEM microwell surfaces for scalable well standardized stem cell aggregation and EB formation. The non-adhesive agarose microwell plates represent an accurate replication of the templates' topography and were used for aggregation of murine induced pluripotent stem cells (iPSCs) and human embryonic stem cells (ESCs). Direct microscopic assessment by time-lapse analysis demonstrated rapid formation of uniformly shaped EBs from murine iPSCs with similar or even more consistent results concerning size distribution and harvesting efficiency compared to the commonly used but time-consuming hanging drop technique. For human ESCs, homogenous aggregates were obtained after single cell inoculation on agarose microwells with efficient differentiation into the cardiac lineage using state-of-the-art protocols for directed differentiation via small molecules. With this soft lithography-based strategy, sufficient and reproducible numbers of stem cell-derived cardiomyocytes necessary for tissue engineering purposes can be realized in a highly controllable manner. Moreover, it might be useful for different cell types in any application that requires scalable and highly standardized aggregation.
authors: Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Hannover Medical School, 30625 Hannover, REBIRTH Cluster of Excellence, Germany.
authors: Julia
authors: J
authors: Dahlmann
authors: George
authors: G
authors: Kensah
authors: Henning
authors: H
authors: Kempf
authors: David
authors: D
authors: Skvorc
authors: Anke
authors: A
authors: Gawol
authors: David A
authors: DA
authors: Elliott
authors: Gerald
authors: G
authors: Dräger
authors: Robert
authors: R
authors: Zweigerdt
authors: Ulrich
authors: U
authors: Martin
authors: Ina
authors: I
authors: Gruh
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D058732:Embryoid Bodies / Q000166:cytology*; D053595:Embryonic Stem Cells / Q000166:cytology*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D051379:Mice; D012685:Sepharose / Q000737:chemistry*; D023822:Tissue Engineering
chemical_list: D012685:Sepharose